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| United States Patent |
5,786,347
|
|
Hesse
, et al.
|
July 28, 1998
|
Vitamin D amine and amide derivatives
Abstract
The invention relates to vitamin D amine and amide derivatives of general
formula
##STR1##
where R represents a hydrogen atom, an aliphatic, cycloaliphatic or
araliphatic group, or an acyl group comprising an aliphatic,
cycloaliphatic, arylaliphatic or aryl group linked to the nitrogen atom by
way of a carbonyl group; R.sup.1 and R.sup.2 are each selected from lower
alkyl and cycloalkyl groups or together with the carbon atom to which they
are attached form a lower cycloalkyl group; R.sup.3 represents a methyl
group having .alpha.- or .beta.-configuration; Y represents a lower
alkylene, alkenylene or alkynylene group optionally substituted by a
hydroxyl, etherified hydroxyl or esterified hydroxyl group; and A.dbd.
represents a cyclohexylidene moiety characteristic of the A-ring of a 1
.alpha.-hydroxylated vitamin D or analogue thereof. Active compounds of
the invention exhibit cell modulating activity and in certain cases may
also have an effect on calcium metabolism. The compounds of the invention
may be prepared by isomerizing a 5,6-trans isomer of formula (I) to a
corresponding 5,6-cis isomer; by hydroxylating a 1-unsubstituted-5,6-trans
analogue of a compound of formula (I) to prepare a 5,6-trans isomer of
formula (I); by reacting a compound containing a precursor for the desired
17-position side chain in one or more stages and with one or more
reactants serving to form the desired side chain; or by reacting a
compound of formula (I) to modify the substitution pattern about the
A.dbd. group.
| Inventors:
|
Hesse; Robert Henry (Winchester, MA);
Setty; Sundara Katugam Srinivasaetty (Cambridge, MA);
Ramgopal; Malathi (Andover, MA)
|
| Assignee:
|
Research Institute for Medicine and Chemistry, Inc. (Cambridge, MA)
|
| Appl. No.:
|
652597 |
| Filed:
|
July 31, 1996 |
| PCT Filed:
|
December 13, 1994
|
| PCT NO:
|
PCT/GB94/02725
|
| 371 Date:
|
July 31, 1996
|
| 102(e) Date:
|
July 31, 1996
|
| PCT PUB.NO.:
|
WO95/16672 |
| PCT PUB. Date:
|
June 22, 1995 |
Foreign Application Priority Data
| Current U.S. Class: |
514/167; 552/653 |
| Intern'l Class: |
A01N 045/00; C07C 401/00 |
| Field of Search: |
552/653
514/167
|
References Cited
| Foreign Patent Documents |
| 93/09093 | May., 1993 | WO.
| |
Primary Examiner: Dees; Jose G.
Assistant Examiner: Pryor; Alton
Attorney, Agent or Firm: Bacon & Thomas
Claims
We claim:
1. A compound of general formula (I)
##STR17##
where R represents a hydrogen atom, an aliphatic, cycloaliphatic or
araliphatic group, or an acyl group comprising an aliphatic,
cycloaliphatic, arylaliphatic or aryl group linked to the nitrogen atom by
way of a carbonyl group;
R.sup.1 and R.sup.2 are each selected from lower alkyl and cycloalkyl
groups or together with the carbon atom to which they are attached form a
lower cycloalkyl group;
R.sup.3 represents a methyl group having .alpha.- or .beta.-configuration;
Y represents a lower alkylene, alkenylene or alkynylene group optionally
substituted by a hydroxyl, etherified hydroxyl or esterified hydroxyl
group; and
A.dbd. represents a cyclohexylidene moiety characteristic of the A-ring of
a 1.alpha.-hydroxylated vitamin D or analogue thereof.
2. A compound as claimed in claim 1 wherein R represents a hydrogen atom, a
lower alkyl or a lower alkanoyl group.
3. A compound as claimed in claim 1 in which R.sup.1 and R.sup.2 are each
selected from methyl, ethyl, propyl and butyl groups.
4. A compound as claimed in claim 1 wherein Y is a straight chain group
containing 3-6 carbon atoms.
5. A compound as claimed in claim 4 wherein Y is selected from
trimethylene, tetramethylene, pentamethylene, hexamethylene,
buta-1,3-dienylene, propynylene, but-1-ynylene and but-2-ynylene.
6. A compound as claimed in claim 1 wherein Y carries a hydroxy, etherified
hydroxy or esterified hydroxy group in a position .alpha.-, .beta.- or
.gamma.- to the group --C(R.sup.1) (R.sup.2).NHR or .alpha.- to any triple
bond present in the group Y.
7. A compound as claimed in claim 1 wherein A.dbd. represents one of the
groups
##STR18##
where R.sup.4 and R.sup.5 are each selected from hydrogen atoms and
O-protecting groups.
8. A compound as claimed in claim 7 wherein R.sup.4 and R.sup.5 represent
etherifying silyl groups.
9. A compound as claimed in claim 7 wherein R.sup.4 and R.sup.5 are
selected from hydrogen atoms and metabolically labile etherifying or
esterifying groups.
10. A compound as claimed in claim 1 wherein A.dbd. represents one of the
groups
##STR19##
11. A 20,20-dimethyl, 20-methylene or 20-spirocyclopropyl analogue of a
compound as claimed in claim 1.
12. A Pharmaceutical composition comprising an active compound as claimed
in claim 1 in admixture with one or more physiologically acceptable
carriers or excipients.
13. A method of treatment of a human or animal subject to promote treatment
and/or prevention of rickets, osteomalacia, osteoporosis,
hypoparathyroidism, hypophosphataemia, hypocalcaenia and/or associated
bone disease, hypocalcaemic tetiary, renal failure, renal osteodystraphy,
biliary cirrhosis, steatorrhea, secondary hypocalcaemia and/or associated
bone disease, wound healing, fertility control, suppression of parathyroid
hormone or management of disorders involving blood clotting or to combat
neoplastic disease, infection, bone disease, autoimmune disease,
host-graft reaction, transplant rejection, inflammatory disease,
neoplasia, hyperplasia, myopathy, enteropathy, spondylitic heart disease,
dermatological disease, hypertension, rheumatoid arthritis, psoriatic
arthritis, secondary hyperparathyroidism, asthma, cognitive impairment or
senile dementia, comprising administration to said subject of an effective
amount of an active compound as claimed in claim 1.
14. A process for the preparation of a compound of general formula (I) as
defined in claim 1 which comprises one or more of:
A) isomerising a 5,6-trans isomer of general formula (I) to a corresponding
5,6-cis isomer, followed if necessary and/or desired by removal of any
O-protecting groups;
B) hydroxylating a 1-unsubstituted-5,6-trans analogue of a compound of
general formula (I) to prepare a 5,6-trans isomer of general formula (I),
followed if necessary and/or desired by isomerisation and/or removal of
any O-protecting group;
C) reacting a compound containing a precursor for the desired 17-position
side chain in one or more stages and with one or more reactants serving to
form the desired side chain, followed if necessary and/or desired by
isomerisation and/or removal of any O-protecting groups; or
D) reacting a compound of formula (I) to modify the substitution pattern
about the A.dbd. group, followed if necessary and/or desired by
isomerisation and/or removal of protecting groups.
15. A compound:
25-amino-1.alpha.,3.beta.-dihydroxy-9,10-secocholesta-5(Z),7,10(19)-triene;
25-amino-1.alpha.,3.beta.-dihydroxy-23,24-bisnor-9,10-secocholesta-5(Z),7,1
0(19)-triene;
25-acetamido-1.alpha.,3.beta.-dihydroxy-9,10-secocholesta-5(Z),7,10(19)-tri
ene;
25-amino-1.alpha.,3.beta.-dihydroxy-20-epi-9,10-secocholesta-5(Z),7,10(19)-
triene;
25-amino-1.alpha.,3.beta.-dihydroxy-24-homo-9,10-secocholesta-5(Z),7,10(19)
-triene;
25-amino-1.alpha.,3.beta.-dihydroxy-9,10-secocholesta-5(Z),7,10(19)-triene;
25-amino-1.alpha.,3.beta.-dihydroxy-9,10-secocholesta-5(E),7,diene;
25-amino-1.alpha.,3.beta.-dihydroxy-9,10-secocholesta-5(Z),7,diene;
25-amino-1.alpha.,3.beta.-dihydroxy-10-spirocyclopropyl-9,10-secocholesta-5
(Z),7,10(19)-triene;
25-amino-1.alpha.,3.beta.-dihydroxy-10-spirocyclopropyl-9,10-secocholesta-5
(E),7,10(19)-triene;
25-amino-1.alpha.,3.beta.-dihydroxy-20-epi-19-nor-9,10-secocholesta-5,7-die
ne;
25-amino-1.alpha.,3.beta.-dihydroxy-24,26,27-tris-homo-9,10-secocholesta-5(
Z),7,10(19)-triene;
25-amino-1.alpha.,3.beta.-dihydroxy-24,26,26,26,27,27,27-heptakis-homo-9,10
-secocholesta-5(Z),7,10(19)-triene;
25-acetamido-1.alpha.,3.beta.-dihydroxy-24-homo-9,10-secocholesta-5(Z),7,10
(19)-triene;
25-amino-1.alpha.,3.beta.-dihydroxy-26,27-bis-homo-9,10-secocholesta-5(Z),7
,10(19)-trien-23-yne;
25-amino-1.alpha.,3.beta.-dihydroxy-20-epi-24,26,27-tris-homo-9,10-secochol
esta-5(Z),7,10(19)-trien-24(24a)-yne;
25-amino-1.alpha.,3.beta.-dihydroxy-24-homo-9,10-secocholesta-5(Z),7,10(19)
-trien-24(24a)-yne;
25-amino-1.alpha.,3.beta.-dihydroxy-20-epi-24,26,27-tris-homo-9,10-secochol
esta-5(Z),7,10(19)-triene;
25-amino-1.alpha.,3.beta.-trihydroxy-26,27-bis-homo-9,10-secocholesta-5(Z),
7,10(19)-trien-23-yne;
25-amino-1.alpha.,3.beta.,23-trihydroxy-24,26,27-tris-homo-9,10-secocholest
a-5(Z),7,10(19)-trien-24(24a)-yne;
25-amino-1.alpha.,3.beta.,22-trihydroxy-9,10-secocholesta-5(Z),7,10(19)-tri
en-23-yne;
25-amino-1.alpha.,3.beta.-dihydroxy-22-methoxy-9,10-secocholesta-5(Z),7,10(
19)-trien-23-yne;
25-amino-1.alpha.,3.beta.-dihydroxy-22-ethoxy-9,10-secocholesta-5(Z),7,10(1
9)-trien-23-yne;
25-amino-1.alpha.,3-dihydroxy-22-propoxy-9,10-secocholesta-5(Z),7,10(19)-tr
ien-23-yne;
N-ethyl-25-amino-1.alpha.,3.beta.-dihydroxy-24-homo-9,10-secocholesta-5(Z),
7,10(19)-triene;
25-benzamido-1.alpha.,3.beta.-dihydroxy-24-homo-9,10-secocholesta-5(Z),7,10
(19)-triene; or
25-amino-1.alpha.,3.beta.-dihydroxy-24,26,27-tris-homo-9,10-secocholesta-5(
Z),7,10(19),22,24(24a)-pentaene.
Description
This invention relates to novel vitamin D analogues, more particularly to
1.alpha.-hydroxy vitamin D.sub.3 analogues having a modified side chain at
the 17-position.
Vitamin D.sub.3, which has the formula
##STR2##
is well known to play a vital role in the metabolism of calcium, by
promoting intestinal absorption of calcium and phosphorus, maintaining
adequate serum levels of calcium and phosphorus and stimulating
mobilisation of calcium from the bone fluid compartment in the presence of
parathyroid hormone.
It was learned more than 20 years ago that the D vitamins undergo
hydroxylation in vivo, hydroxylation at the 25-position occurring in the
liver and hydroxylation at the 1.alpha.-position occurring in the kidney,
the resulting 1.alpha.,25-dihydroxy metabolite being the biologically
active material. This discovery led to the synthesis of many analogues of
vitamin D, evaluation of which indicated that hydroxyl groups at the
1.alpha.-position a and at either the 24R- or the 25-position were
essential for a compound or metabolite thereof to exhibit a substantial
effect on calcium metabolism. While, as indicated above, such hydroxyl
groups will normally ultimately be introduced in vivo, hydroxylation at
the 24R- or 25-position occurring rather more readily than at the
1.alpha.-position, the use of vitamin D analogues already so hydroxylated
has proved of substantial advantage by virtue of their enhanced levels of
activity and their rapidity of action and subsequent elimination from the
body. It will be appreciated that 1.alpha.-hydroxylated vitamin D
derivatives are of especial benefit to patients suffering from renal
failure.
Examples of hydroxylated vitamin D analogues in current use include the
natural metabolite 1.alpha.,25-dihydroxy vitamin D.sub.3 and
1.alpha.-hydroxy vitamin D.sub.3 (which is readily 25-hydroxylated in
vivo). Other reportedly promising compounds include 1
.alpha.,24R-dihydroxy vitamin D.sub.3, D.sub.2 analogues of the above
compounds and 1.alpha.,25-dihydroxy analogues carrying fluorine atoms at
the 24-, 26- and/or 27- positions (see De Luca and Schnoes, Ann. Rev.
Biochem. (1983), 52, pp 411-439 and De Luca et al., Top. Curr. Chem.
(1979), 83, pp 1-65).
More recently it has been learned that the natural metabolite
1.alpha.,25-dihydroxy vitamin D.sub.3 has additional effects on cellular
metabolism. These cell modulating effects include stimulation of cell
maturation and differentiation (Tanaka et al., Biochem. J. (1982), 204, pp
713-719; Amento et al., J. Clin. Invest, (1984),73, pp 731-739; Colston et
al., Endocrinology (1981), 108, pp 1083-1086; Abe et al., Proc. Nat, Acad.
Sci. (1981), 78, pp 4990-4994) and immunosuppressive effects (e.g.
inhibition of interleukin II production) (Rigby, Immunology Today (1988),
9, pp 54-58).
Still more recently, an immunopotentiating effect of 1.alpha.,25-dihydroxy
vitamin D.sub.3 has been observed, the compound having been found to
stimulate the production of bactericidal oxygen metabolites and the
chemotactic response of leukocytes (see, for example, Cohen et al., J.
Immunol. (1986), 136, pp 1049-1053). It is well known that leukocytes play
a major role in the body's defence against various infections (see, for
example, Roitt, Brostoff and Male, "Immunology" 2nd Ed. (1989), C. V.
Mosby, St. Louis, sec 16.10-16.13 and 17.4-17.5), e.g. by adhering to and
engulfing invading organisms (chemotactic response) and/or by producing
superoxides and/or other toxic oxygen metabolites. It is known that this
response may also be stimulated by mitogens such as the co-carcinogenic
phorbal esters and .gamma.-interferon, which are structurally quite
different from vitamin D analogues.
By virtue of these effects on cellular metabolism, 1.alpha.,25-dihydroxy
vitamin D.sub.3 in principle has therapeutic potential in such diverse
areas as treatment of psoriasis, inflammatory and autoimmune diseases,
neoplasias and hyperplasias, as an adjunct in the chemotherapy of
infections (inter alia bacterial, viral and fungal), and in other
therapeutic modalities in which mononuclear phagocytes are involved.
1.alpha.,25-dihydroxy vitamin D.sub.3 and 1.alpha.-hydroxy vitamin D.sub.3
have also been proposed for use in the treatment of hypertension (Lind et
al., Acta Med. Scand. (1987), 222, pp 423-427) and diabetes mellitus
(Inomata et al., Bone Mineral (1986), 1, pp 187-192), and it has been
suggested that 1.alpha.,25-dihydroxy vitamin D.sub.3 may promote hair
growth (Lancet, 4 Mar. 1989, p 478) and may be useful in the treatment of
acne (Malloy et al., Tricontinental Meeting for Investigative Dermatology,
Washington, 1989).
The potent effects of 1.alpha.,25-dihydroxy vitamin D.sub.3 and
1.alpha.-hydroxy vitamin D.sub.3 on calcium metabolism will, however,
normally preclude such uses, since dosages at a level sufficient to elicit
a desired cell modulating, immunosuppressive or immunopotentiating effect
tend to lead to unacceptable hypercalcaemia. This has led to attempts to
synthesize new analogues having reduced effects on calcium metabolism but
which still exhibit the desired effects on cellular metabolism.
There have been reports of new analogues which exhibit, to at least a
moderate degree, this desired separation of activity. Thus the compound
MC-903 (calcipotriol), which is a 22,23-unsaturated 1.alpha.,24R-dihydroxy
vitamin D.sub.3 analogue carrying a cyclopropyl group at the 24-position
instead of the usual C.sub.25-C.sub.27 configuration of the cholestane
side chain, and which is now used for the treatment of psoriasis, is
reported to exhibit an effect on cell maturation comparable in magnitude
to 1.alpha.,25-dihydroxy vitamin D.sub.3, while exhibiting a smaller
hypercalcaemic effect (Calverley, Tetrahedron (1987), 43, pp 4609-4619;
and Holick, Arch, Dermatol. (1989), 125, pp 1692-1696). Similar claims
have been made for analogues of 1.alpha.,25-dihydroxy vitamin D.sub.3,
e.g. the 22-oxa (Abe et al., Endocrinology (1989), 124, pp 2645-2647), the
24- and the 26- homo (Ostrem et al., J. Biol, Chem. (1987), 262, pp
14164-14171), the 16-dehydro- 23,24-ethynyl (Zhou et al., Blood (1989),
74, pp 82-93) and the 19-nor-1-dihydro (Perlman et al., Tetrahedron Lett.
(1990), pp 1823-1824).
Other analogues of 1.alpha.,25-dihydroxy vitamin D.sub.3 which have been
studied with the aim of achieving enhanced separation of
differentiation-inducing activity and hypercalcaemic effect include
23-oxa, 23-thia and 23-aza derivatives (Kubodera et al., Chem. Pharm.
Bull. (1991), 39, pp 3221-3224), 22-oxa analogues bearing side chains of
different sizes (Kubodera et al., Chem. Pharm. Bull. (1992), 40, pp
1494-1499), and 20-epi analogues (Binderup et al., Biochemical
Pharmacology (1991), 42, pp 1569-1575).
It does not appear possible to deduce from these disclosures either which
compounds will exhibit cell modulating activity(or the level of any such
activity) or to determine factors which lead to a separation of activities
as regards cell modulation and calcium metabolism. Thus, for example, it
has been observed that there are no strict relationships between
differentiation-inducing activity and side chain length or hydrophilicity.
The majority of results suggest that the presence of a hydroxyl group
towards the end of a cholestane-type side chain or homologue thereof is
necessary for compounds to show significant cell modulating activity.
However, the findings of Ostrem et al. (op. cit.) indicate that analogues
having only a short, unsubstituted 17-position side chain (e.g. isopropyl
or sec-butyl, as in homo- or bis-homo-pregnanes) exhibit quite
substantial-differentiation-inducing activity and are more potent than
corresponding short side chain compounds bearing a side chain hydroxyl
group.
A number of the proposed analogues appear to show cell modulating activity
at a similar level to that of 1.alpha.,25-dihydroxy vitamin D.sub.3, but
also appear still to show appreciable effects on calcium metabolism, such
activity being attenuated by at most two orders of magnitude relative to
that of 1.alpha.,25-dihydroxy vitamin D.sub.3. Moreover, it now appears in
the case of many, if not all, of the new analogues described above as
exhibiting separation of calcium and cellular metabolic effects, including
MC-903, that the attenuated calcium effect may be due merely to more rapid
metabolism of the vitamin reducing the amount of the circulating drug (see
e.g. Bouillon et al., J. Bone Miner. Res. (1991), 6, p 1051 and Dusso et
al., Endocrinology (1991), 128, p 1687). This may similarly reduce the
cell modulating effect in vivo so that one may require larger systemic
dosages than are suggested by in vitro test results.
Use of such analogues may therefore give rise to cumulative toxicity
problems if the compounds are used in long term therapy, particularly
where systemic application is required, e.g. for treatment of inflammatory
and autoimmune diseases, neoplasias and hyperplasias, or in oral therapy
for treatment of psoriasis, and there is thus a continuing need in such
areas of therapy for vitamin D-like compounds which exhibit potent cell
modulating activity coupled with a reduced effect on calcium metabolism.
There may also be circumstances in which a particular balance of cell
modulating and calcium metabolising properties is desired. This may be the
case in, for example, the treatment of osteoporosis. The present invention
is based on the finding that 1.alpha.-hydroxy vitamin D derivatives in
which the 17-position side chain carries certain amine or amide functions
may exhibit useful biological activity; this is most surprising in that
the invention includes is compounds which lack a side chain hydroxyl
group; such a group has hitherto normally been thought desirable in order
to promote calcaemic and/or cell modulating activity. Furthermore, as
noted by Kubodera et al. (op. cit., 1991), introduction of a nitrogen atom
into the 17-position side chain of 1.alpha.,25-dihydroxy vitamin D.sub.3
to replace a methylene group thereof appears to be deactivating as regards
differentiation-inducing activity. One might therefore expect replacement
of the activity-promoting side chain hydroxyl group of a vitamin D
analogue by an amine or amide function to be even more deactivating,
especially since amine groups are very significantly more basic than
oxygen functions such as hydroxyl groups, and are protonated at
physiological pH.
Additionally, as is described in greater detail hereinafter, it has been
found that by appropriate selection of e.g. the size of the carbon chain
of the 17-position side chain of the compounds according to the invention
it is possible to influence their activity by enhancing either cell
modulating properties or activity as regards calcium metabolism and bone
calcium mobilisation, thereby making possible the preparation of compounds
with particular activity profiles suited to particular therapeutic
applications.
Thus according to one aspect of the present invention there are provided
compounds of formula (I)
##STR3##
(where R represents a hydrogen atom, an aliphatic, cycloaliphatic,
araliphatic or aryl organic group, or an acyl group comprising such an
organic group linked to the nitrogen atom by way of a carbonyl group;
R.sup.1 and R.sup.2, which may be the same or different, each represent a
lower alkyl or cycloalkyl group or together with the carbon atom to which
they are attached form a lower cycloalkyl group; R.sup.3 represents a
methyl group having .alpha.a-or .beta.-configuration; Y represents a lower
alkylene, alkenylene or alkynylene group optionally substituted by a
hydroxyl, etherified hydroxyl or esterified hydroxyl group; and A.dbd.
represents a cyclohexylidene moiety characteristic of the A-ring of a
1.alpha.-hydroxylated vitamin D or analogue thereof.
Where either of R.sup.1 and R.sup.2 represent lower alkyl groups these may,
for example, be C.sub.1-6 alkyl groups such as methyl, ethyl, propyl and
butyl groups. Lower cycloalkyl groups R.sup.1 and R.sup.2 may, for
example, contain 3-8 carbon atoms, e.g. as in cyclopropyl, cyclopentyl and
cyclohexyl groups.
Where R represents an aliphatic or cycloaliphatic group this may, for
example, be a lower alkyl or lower cycloalkyl group, e.g. as described for
R.sup.1 and R.sup.2. Araliphatic groups R may, for example, include
C.sub.6-12 carbocyclic aryl C.sub.1-4 alkyl groups such as benzyl or
phenethyl; aryl groups may, for example, include C.sub.6-12 carbocyclic
aryl groups such as phenyl or naphthyl. Where R represents an acyl group
this may, for example, be a lower (e.g. C.sub.1-6) alkanoyl group such as
formyl, acetyl or propionyl; a C.sub.6-12 carbocyclic aryl C.sub.2-5
alkanoyl group such as phenylacetyl; or a C.sub.7-13 carbocyclic aroyl
group such as benzoyl. The group R may optionally carry one or more
substituents, for example selected from halo (e.g. chloro or bromo), lower
(e.g. C.sub.1-4) alkyl such as methyl, lower alkoxy (e.g. methoxy), lower
alkanoyl (e.g. acetyl), lower alkylamino (e.g. methylamino), di(lower
alkyl)amino (e.g. dimethylamino), nitro, carbamoyl and lower alkanoylamino
(e.g. acetamido).
Lower alkylene, alkenylene or alkynylene groups represented by Y may, for
example, contain up to 7 carbon atoms and up to 3 multiple bonds. Y may
advantageously be a straight chained group, e.g. containing 3-6 carbon
atoms, for example as in trimethylene, tetramethylene, pentamethylene,
hexamethylene, buta-1,3-dienylene, propynylene, but-1-ynylene or
but-2-ynylene.
Where Y is substituted by a hydroxyl, etherified hydroxyl or esterified
hydroxyl group, this substituent may advantageously be positioned
.alpha.-, .beta.- or .gamma.- to the group --C(R.sup.1) (R.sup.2) .NHR or
.alpha.- to any triple bond present in the group Y. Etherified hydroxyl
groups which may be present inclkude lower (e.g. C.sub.1-6) alkyl groups
optionally interrupted by one or more oxygen atoms (e.g. methyl,
methoxymethyl or methoxyethoxymethyl), and cyclic groups such as
tetrahydropyranyl. Esterified hydroxyl groups which may be present include
lower (e.g. C.sub.1-6) alkanoyl such as acetyl, propionyl, isobutyryl or
pivaloyl; lower alkenoyl (e.g. allylcarbonyl); aroyl (e.g.
p-nitrobenzoyl); lower alkoxycarbonyl (e.g. t-butoxycarbonyl); lower
haloalkoxycarbonyl (e.g. 2,2,2-trichloroethoxycarbonyl or
1,1,1-trichloro-2-methyl-2-propoxycarbonyl); aralkyloxycarbonyl (e.g.
benzyloxycarbonyl or p-nitrobenzyloxycarbonyl); and lower
alkenyloxycarbonyl (e.g. allyloxycarbonyl). It will be appreciated that it
may be advantageous to select etherifying or esterifying groups which are
metabolically labile in vivo.
Where R.sup.3 in formula (I) is a methyl group in the .alpha.-configuration
the compounds have the 20R configuration characteristic of natural vitamin
D derivatives; where R.sup.1 is in the .beta.-configuration the compounds
have the 20S configuration of epi-vitamin D derivatives. It will be
appreciated that the invention also embraces mixtures of the two isomers.
The cyclohexylidene ring represented by A.dbd. will normally carry hydroxyl
groups or protected derivatives thereof at the 1.alpha.- and
3.beta.-positions, and may carry further substituents, e.g. which tend to
enhance calcaemic or antiproliferative activity and/or stimulate
differentiation. A.dbd. may thus, for example, be represented by the
formula (A-1)
##STR4##
where R.sup.4 and R.sup.5, which may be the same or different, each
represent a hydrogen atom or an 0-protecting group, and R.sup.6 and
R.sup.7, which may the same or different, are selected from hydrogen atoms
and appropriate mono- or di-valent substituting groups.
Where R.sup.4 and R.sup.5 represent O-protecting groups these may, for
example, be cleavable O-protecting groups such as are commonly known in
the art. Suitable groups include etherifying groups such as silyl groups
(e.g. tri (lower alkyl) silyl groups such as trimethylsilyl,
triethylsilyl, triisopropylsilyl or t-butyldimethylsilyl; tri (aryl) silyl
groups such as triphenylsilyl; and mixed alkyl-arylsilyl groups); lower
(e.g. C.sub.1-6) alkyl groups optionally interrupted by an oxygen atom,
such as methyl, methoxymethyl or methoxyethoxymethyl; and cyclic groups
such as tetrahydropyranyl. Esterifying O-protecting groups include lower
(e.g. C.sub.1-6) alkanoyl such as acetyl, propionyl, isobutyryl or
pivaloyl; aroyl (e.g. containing 7-15 carbon atoms) such as benzoyl or
4-phenylazobenzoyl; lower alkane sulphonyl such as (optionally
halogenated) methane sulphonyl; and arene sulphonyl such as p-toluene
sulphonyl.
O-protected derivatives are useful as intermediates in the preparation of
active 1.alpha.,3.beta.-diols of formula (I) where R.sup.4 and R.sup.5
represent hydrogen atoms. Additionally, where the O-protecting groups are
metabolically labile in vivo, such ethers and esters of formula (I) may be
useful directly in therapy.
At least one of R.sup.6 and R.sup.7 is advantageously a hydrogen atom.
Substituents which may be present as the. other of R.sup.6 and R.sup.7
include, for example, methylene, methyl and ethylene (so as to form a
spiro-linked cyclopropyl group with the attached carbon atom).
Representative A.dbd. groups falling within the above formula A-1) include
the following:
##STR5##
It will be appreciated that compounds containing groups (A-2) and (A-3) are
respectively 5,6-cis (i.e. 5 Z) and 5,6-trans (i.e. 5 E) isomers of
vitamin D analogues. Compounds containing groups (A-4) and (A-5) are
similarly 5,6-cis and 5,6-trans isomers respectively of 10,19-dihydro
vitamin D analogues, and compounds containing group (A-8) are 19-nor
vitamin D analogues.
5,6-Trans isomers according to the invention are principally of interest as
intermediates in the preparation of corresponding 5,6-cis isomers, e.g. as
described in greater detail hereinafter. However, 5,6-trans isomers in
which R.sup.4 and R.sup.5 are hydrogen atoms or metabolically labile
groups will often exhibit biological activity, e.g. at about one order of
magnitude less than corresponding 5,6-cis isomers, and may thus be useful
in therapy.
Active compounds of formula (I) in which Y is, for example, a group
containing up to 3 carbon atoms such as trimethylene and R.sup.1 and
R.sup.2 are, for example, lower alkyl groups such as methyl or ethyl may
exhibit similar activity to known 1.alpha.-hydroxy vitamin D derivatives
such as 1.alpha.,25-dihydroxy vitamin D.sub.3. Thus, for example, such
compounds may exhibit a significant effect on calcium metabolism, e.g. by
stimulating intestinal calcium transport, bone calcium mobilisation and
bone formation. These compounds may therefore have applications in, for
example, treatment and/or prevention of disorders such as rickets and
osteomalacia, osterporosis, hypoparathyroidism, hypophosphateamia,
hypocalcaemia and/or associated bone disease, hypocalcaemic tetany, renal
failure and disorders such as renal osteodystrophy, biliary cirrhosis and
steatorrhea, and secondary hypocalcaemia and/or bone disease arising from
disfunction of the liver, kidneys or gastrointestinal tract or resulting
from treatment with dilantin, barbiturates such as phenylbarbitone and
related drugs; they may be particularly useful in treating disorders which
are refractory to natural compounds such as vitamin D.sub.3.
The presence of an amino group in compounds of formula (I) in which R
represents hydrogen or a lower alkyl group may enhance the bioavailability
of the compounds relative to vitamin D derivatives containing a side chain
hydroxyl group, for example by limiting sequestration in liposomes, and
may also permit the compounds readily to be formulated as aqueous
compositions, a highly advantageous property compared to conventional
oil-soluble vitamin D derivatives.
The above-described compounds of formula (I) also exhibit cell modulating
activity, e.g. as evidenced by eliciting cell differentiation and
maturation, inhibiting proliferation and/or by activating monocytes (e.g.
as estimated by the method of Styrt et al., Blood (1986), 67, pp 334-342),
but their calcaemic effects may be too pronounced to permit e.g. mid- or
long-term use in respect of their cellular metabolic effects. The higher
homologues of these compounds, however, e.g. compounds (I) in which Y
contains 4-7 carbon atoms and/or R.sup.1 and/or R.sup.2 each contain 2 or
more carbon atoms, may tend to exhibit a reduced effect on calcium
metabolism, e.g. as evidenced by low effects on serum calcium and
phosphorus levels in rats, and may accordingly exhibit an advantageous
therapeutic ratio of cell modulating to calcaemic activity.
The cell modulating activity of such active compounds according to the
invention, combined with a substantial lack of calcaemic effect, render
them of interest (both alone and as adjuncts) in the management of
neoplastic disease, particularly myelogenous leukemias, and suggest their
use as agents to promote wound healing. They may also be used either alone
or as adjuncts in the chemotherapy of infection and in all other
therapeutic modalities in which mononuclear phagocytes are involved, for
example in treatment of bone disease (e.g. osteoporosis, osteopenia and
osteodystrophy as in rickets or renal osteodystrophy), autoimmune
diseases, host-graft reaction, transplant rejection, inflammatory diseases
(including modulation of immunoinflammatory reactions), neoplasias and
hyperplasias, myopathy, enteropathy and spondylitic heart disease. Such
active compounds according to the invention may also be useful in
suppression of parathyroid hormone (e.g. as in serum calcium homeostasis),
in treatment of dermatological diseases (for example including acne,
alopecia, eczema, pruritus, psoriasis and skin aging, including
photoaging), hypertension, rheumatoid arthritis, psoriatic arthritis,
secondary hyperparathyrodism, asthma, cognitive impairment and senile
dementia (including Alzheimer's disease), in fertility control in both
human and animal subjects, and in management of disorders involving blood
clotting, e.g. by dissolution of existing clots and/or prevention of
clotting. The invention embraces use of these compounds in the therapy or
prophylaxis of such conditions and in the manufacture of medicaments for
such treatment or prophylaxis.
We believe that the active 20R isomers of such compounds of formula (I) may
be preferred for treatment of infections, e.g. in combination therapy,
whereas the active 20S epi-isomers may be preferred for applications
involving an immunosuppressive effect, e.g. in treatment of autoimmune and
inflammatory diseases, rheumatoid arthritis, asthma etc. This view is
supported by, for example, the work of Binderup et al. concerning
20-epi-vitamin D.sub.3 analogues reported in Biochemical Pharmacology
(1991), 42(8), pp 1569-1575.
It will be appreciated that it may be preferred to select lower homologues
according to formula (I) for treatment of e.g. defects of calcium
metabolism and to select higher homologues specifically for their cell
modulating activity, e.g. in treatment of hyperplasias such as psoriasis.
However, both types of activity may be useful in, for example, treatment
of bone disease, and particular homologues may therefore be chosen by
selection of appropriate meanings for Y, R.sup.1 and R.sup.2 in order to
give a desired balance of activities for such purposes.
It has been reported (Neef et al., 9th Workshop on Vitamin D (1994) that in
the case of vitamin D compounds having conventional terminally
hydroxylated 17-position side chains (including side chains containing a
heteroatom at the 23-position), analogues having 20,20-dimethyl,
20-methylene or 20-spirocyclopropyl groups may exhibit useful biological
activity, typically resembling that of the corresponding 20R
methyl-substituted isomer rather than the corresponding 20S epi-isomer.
The present invention embraces analogues of the above-defined compounds of
formula (I) wherein R.sup.3 is selected from dimethyl, methylene and
spirocyclopropyl groups.
Active compounds according to the invention may be formulated for
administration by any convenient route, e.g. orally (including
sublingually), parenterally, rectally or by inhalation; pharmaceutical
compositions so formulated comprise a feature of the invention.
Orally administrable compositions may, if desired, contain one or more
physiologically compatible carriers and/or excipients and may be solid or
liquid. The compositions may take any convenient form including, for
example, tablets, coated tablets, capsules, lozenges, aqueous or oily
suspensions, solutions, emulsions, syrups, elixirs and dry products
suitable for reconstitution with water or another suitable liquid vehicle
before use. The compositions may advantageously be prepared in dosage unit
form. Tablets and capsules according to the invention may, if desired,
contain conventional ingredients such as binding agents, for example
syrup, acacia, gelatin, sorbitol, tragacanth or polyvinyl-pyrollidone;
fillers, for example lactose, sugar, maize-starch, calcium phosphate,
sorbitol or glycine; lubricants, for example magnesium stearate, talc,
polyethylene glycol or silica; disintegrants, for example potato starch;
or acceptable wetting agents such as sodium lauryl sulphate. Tablets may
be coated according to methods well known in the art.
Liquid compositions may contain conventional additives such as suspending
agents, for example sorbitol syrup, methyl cellulose, glucose/sugar syrup,
gelatin, hydroxymethylcellulose, carboxymethylcellulose, aluminium
stearate gel or hydrogenated edible fats; emulsifying agents, for example
lecithin, sorbitan monooleate or acacia; non-aqueous vehicles, which may
include edible oils, for example vegetable oils such as arachis oil,
almond oil, fractionated coconut oil, fish-liver oils, oily esters such as
polysorbate 80, propylene glycol, or ethyl alcohol; and preservatives, for
example methyl or propyl p-hydroxybenzoates or sorbic acid. Liquid
compositions may conveniently be encapsulated in, for example, gelatin to
give a product in dosage unit form.
Compositions for parenteral administration may be formulated using an
injectable liquid carrier such as sterile pyrogen-free water, sterile
peroxide-free ethyl oleate, dehydrated alcohol or propylene glycol or a
dehydrated alcohol/propylene glycol mixture, and may be injected
intravenously, intraperitoneally or intramuscularly.
Compositions for rectal administration may be formulated using a
conventional suppository base such as cocoa butter or another glyceride.
Compositions for topical administration include ointments, creams, gels,
lotions, shampoos, paints, powders (including spray powders), pessaries,
tampons, sprays, dips, aerosols, pour-ons and drops. The active ingredient
may, for example, be formulated in a hydrophilic or hydrophobic base as
appropriate.
Compositions for administration by inhalation are conveniently formulated
for self-propelled delivery, e.g. in metered dose form, for example as a
suspension in a propellant such as a halogenated hydrocarbon filled into
an aerosol container provided with a metering dispense valve.
It may be advantageous to incorporate an antioxidant, for example ascorbic
acid, butylated hydroxyanisole or hydroquinone in the compositions of the
invention to enhance their storage life.
Where any of the above compositions are prepared in dosage unit form these
may for example contain 0.1-500 .mu.g, e.g. 0.2-100 .mu.g, of active
compound according to the invention per unit dosage form. The compositions
may if desired incorporate one or more further active ingredients.
A suitable daily dose of an active compound according to the invention may
for example be in the range 0.2-1000 .mu.g, e.g. 0.4-200 .mu.g, per day,
depending on factors such as the severity of the condition being treated
and the age, weight and condition of the subject.
Compounds according to the invention may be prepared by any convenient
method, for example one of the following:
A) 5,6-Cis compounds of formula (I) may be prepared by isomerisation of a
corresponding 5,6-trans compound, followed if necessary and/or desired by
removal of any O-protecting groups. Isomerisation may be effected by, for
example, treatment with iodine, with a disulphide or diselenide, or by
irradiation with ultraviolet light, preferably in the presence of a
triplet sensitiser.
B) 5,6-Trans compounds of formula (I) may be prepared by hydroxylating a
corresponding 1-unsubstituted-5,6-trans compound, e.g. a compound (I)
having an A.dbd. group of the formula
##STR6##
(where R.sup.4 is hydrogen or an O-protecting group). Such hydroxylation
may be effected using a selenite ester (which may be generated in situ by
reaction of selenium dioxide or selenous acid and an alcohol), e.g. as
described in GB-A-2038834, or using selenous acid at a pH in the range
3-9, e.g. as described in GB-A-2108506; the contents of both these
specifications are incorporated herein by reference. The
1-unsubstituted-5,6-trans compound may, if desired, be prepared by
isomerisation of the corresponding 5,6-cis vitamin in situ under the
conditions of the hydroxylation reaction, which may be followed by
isomerisation and/or removal of O-protecting groups as necessary and/or
desired.
C) By reaction of a compound containing a precursor for the desired
17-position side chain in one or more stages and with one or more
reactants serving to form the desired side chain, followed if necessary
and/or desired by isomerisation and/or removal of O-protecting groups.
Thus, for example, in order to prepare a compound (I) in which R.sup.1 and
R.sup.2 are the same, a compound of general formula (II)
##STR7##
(where R.sup.3, Y and A.dbd. are as hereinbefore defined, A.dbd.
preferably being one of the groups (A-2)-(A-8) in O-protected form) may be
reacted with an organo-cerium reagent, e.g. prepared in situ from cerous
chloride and an appropriate organometallic compound, e.g. an
alkyl/cycloalkyl lithium compound of formula R.sup.1 Li (where R.sup.1 is
as hereinbefore defined), for example as described by Ciganek (J. Ora.
Chem. (1992), 5, pp 4521-4527).
Compounds of formula (I) in which R.sup.1 and R.sup.2 are different may,
for example, be prepared by reacting a thio-oxime of formula (III)
##STR8##
(where R.sup.1, R.sup.3, Y and A.dbd. are as hereinbefore defined and
R.sup.8 is an aromatic group, e.g. a carbocyclic aryl group such as
phenyl) with an appropriate organometallic compound, for example an
alkyl/cycloalkyl lithium compound of formula R.sup.2 Li (where R.sup.2 is
as hereinbefore defined), and reducing the thus-obtained compound of
formula (IV)
##STR9##
(where R.sup.1, R.sup.2, R.sup.3.sub.1 R.sup.8, Y and A.dbd. are as
hereinbefore defined), e.g. using a metal hydride reducing agent such as
sodium borohyride or an inorganic or organic sulphur compound such as
hydrogen sulphide, sodium sulphide or a thiol (e.g. a lower alkyl
mercaptan such as methanethiol) to remove the R.sup.8.S group and yield a
corresponding compound of formula (I) in which R is a hydrogen atom (see
J. Ora, Chem. (1977), 42, pp 398-399).
Compounds of formula (I) in which R represents a lower alkanoyl, aralkanoyl
or aroyl group may be prepared by acylation of a corresponding compound
(I) in which R is hydrogen, for example by reaction with an appropriate
acyl halide or acid anhydride or with an appropriate acid in the presence
of a coupling agent such as N,N'-carbonyl-diimidazole or
dicyclohexylcarbodiimide. It will be appreciated that any hydroxyl groups
present elsewhere in the molecule, e.g. as substituents of the A.dbd. or Y
groups, should desirably be in O-protected form during such acylation
reactions.
Compounds of formula (I) in which R represents a lower alkyl group may, for
example, be prepared by reducing a corresponding compound (I) in which R
is a lower alkanoyl group, e.g. using a metal hydride reducing agent such
as lithium aluminium hydride. Alternatively a compound (I) in which R
represents a hydrogen atom may be subjected to direct alkylation, e.g. by
reaction with an alkyl halide, or to reductive amination, e.g. by reaction
with an appropriate aldehyde and a reducing agent such as sodium
cyanoborohydride.
Compounds of formula (I) in which Y is an alkynylene group may, for
example, be prepared by reaction of a compound of formula (V)
##STR10##
(where R.sup.3 and A.dbd. are as hereinbefore defined; Y.sup.a is an
alkylene group, e.g. containing 1-4 carbon atoms; and L represents a
leaving group, for example a sulphonate ester group, e.g. lower alkyl
sulphonyloxy such as mesyloxy, lower fluoroalkyl sulphonyloxy such as
trifluoromethanesulphonyloxy or aryl sulphonyloxy such as tosyloxy, or a
halogen atom such as chlorine, bromine or iodine), with a metallated
derivative (e.g. the lithio derivative) of an alkyne of formula (VI)
##STR11##
(where R, R.sup.1 and R.sup.2 are as hereinbefore defined and n is 0 or an
integer, e.g. in the range 1-3).
The thus obtained compound (I) in which Y is the group
--Y.sup.a --C.tbd.C--(CH.sub.2).sub.n.sup.--
(wherein Y.sup.a and n are as hereinbefore defined) may if desired be
hydrogenated to convert the triple bond either to a double bond (e.g.
using Lindlar catalyst) or to a single bond (e.g. using a noble metal
catalyst such as platinum, palladium or homogeneous rhodium or ruthenium).
During such hydrogenations the 5,7-diene or 5,7,10(19)-triene system of
the compound (I) is preferably protected by formation of a Diels Alder
adduct by reaction with a dienophile, e.g. as described in GB-A-2114570
(the contents of which are incorporated herein by reference); preferred
dienophiles include diacylazo compounds such as phthalazine diones and
phenyl triazoline diones. The Diels Alder adduct may be removed, e.g. by
ozonolysis or other oxidative techniques after the hydrogenation.
Compounds of formula (I) in which Y is an alkynylene group carrying a
hydroxyl group .alpha. to the triple bond may, for example, be prepared by
reaction of a compound of formula (VII)
##STR12##
(where R.sup.3 and A.dbd. are as hereinbefore defined and Y.sup.b is a
valence bond or an alkylene group, e.g. containing 1-4 carbon atoms) with
a metallated derivative of an alkyne of formula (VI), so as to form a
compound (I) in which Y is a group
--Y.sup.b --CH (OH) --C.tbd.--C--(CH.sub.2).sub.n --
(wherein Y.sup.b and n are as hereinbefore defined).
Compounds of the formula (VI) may be prepared by subjecting a compound of
formula (VIII)
CH.sub.3 (CH.sub.2).sub.n C.tbd.C--C(R.sup.1)(R.sup.2)OH
(where n, R.sup.1 and R.sup.2 are as hereinbefore defined) to a Ritter
reaction with a compound of formula R.sup.a CN (where R.sup.a represents a
hydrogen atom or an appropriate organic group) in the presence of a strong
acid, e.g. a mineral acid such as sulphuric acid, thereby leading to
formation of a compound (I) in which R represents a group R.sup.a.CO--.
This group may be removed by hydrolysis to yield a compound (I) in which R
represents a hydrogen atom or may be reduced, e.g. as hereinbefore
described, to yield a compound (I) in which R represents a group
R.sup.a.CH.sub.2 --. Alternatively the hydroxy group of the tertiary
carbinol may be displaced by an azido group, e.g. by reaction with
hydrazoic acid in the presence of a strong acid, and the azido group
reduced to yield a compound (I) in which R represents hydrogen. The
internal alkyne may then be isomerized to the terminal position by
treatment with the potassium salt of 1,3-propanediamine in
1,3-propanediamine as solvent ("acetylene zipper").
Compounds of formula (II) may, for example, themselves be prepared by
reaction of a compound of formula (V) as defined above with, as
appropriate, (i) a source of cyanide ion (e.g. an alkali metal cyanide
such as sodium or potassium cyanide), (ii) a metallated acetonitrile
derivative (e.g. the lithio derivative), or (iii) acrylonitrile,
preferably where L is an iodine atom (e.g. by ultrasound-induced
chromium-mediated conjugate addition as described by Mourino et al. in J
Org. Chem, (1993), 58, pp 118-123).
Compounds (II) in which the 17-position side chain terminates in the group
--CH:CH.CN may, for example, be prepared from an aldehyde of formula (VI)
as defined above by means of a Wittig reaction with an ylid of formula
(R.sup.9).sub.3 P:CH.CN (where each R.sup.9 represents an organic group,
e.g. a carbocyclic aryl group such as phenyl) or with a corresponding
phosphonate or silyl equivalent.
Compounds of formula (III) may, for example, themselves be prepared by
reacting a ketone of formula (X)
##STR13##
(where R.sup.1, R.sup.3, Y and A.dbd. are as hereinbefore defined) with an
S-substituted thiolamine of formula R.sup.8.S.NH (where R.sup.8 is as
hereinbefore defined). Such compounds of formula (IX) may be prepared
from, for example, an acid of formula (X)
##STR14##
(where R.sup.3, Y and A.dbd. are as hereinbefore defined), e.g. by
formation of a corresponding acid halide such as the chloride and reaction
with an organometallic compound R.sup.1 MX (where R.sup.1 is as
hereinbefore defined; M represents a divalent metal such as copper, zinc
or cadmium; and X represents e.g. a halogen atom). Alternatively one may
prepare compounds (IX) by reacting a compound of formula (V) above with
e.g. (i) an .alpha.-metallated derivative such as a lithio derivative of a
ketone of formula CH.sub.3.CO.R.sup.1 (where R.sup.1 is as hereinbefore
defined) or with a corresponding enol, or (ii), preferably where L is an
iodine atom, a vinyl ketone of formula CH.sub.2 :CH.CO.R.sup.1 (where
R.sup.1 is as hereinbefore defined), e.g. by ultrasound-induced
chromium-mediated conjugate addition as described by Mourino et al. (op.
cit.).
Compounds (X) and esters thereof may also be used to prepare compounds of
formula (II) by reaction with ammonia or a metallated derivative thereof,
e.g. an alkali metal amide such as lithium amide, to form a corresponding
carboxamide which may be converted to a nitrile (II) by mild dehydration,
e.g. using tosyl chloride, phosphorus oxychloride in the presence of a
base such as pyridine, or trifluoracetic anhydride in the presence of an
excess of a base such as pyridine.
Compounds (II) in which Y is .alpha.-substituted by a hydroxyl group are
conveniently obtained by cyanohydrin formation, for example by reaction of
a compound (VII) with hydrogen cyanide. Compounds (II) in which Y is
.beta.-substituted by a hydroxyl group may be prepared directly by
reaction of a compound (VII) with a metallated (e.g. lithated) derivative
of acetonitrile; they may also be prepared indirectly by reaction with a
metallated derivative of an ester of acetic acid, followed by conversion
of the ester group to a carboxamide group and then to a nitrile group,
e.g. as described above.
In general compounds (I) and starting materials therefor in which Y is
substituted by a hydroxyl group may be converted to corresponding ether
and ester derivatives by standard methods such as are well known in the
art. Thus, for example, etherification may be effected by reaction with an
appropriate organic halide (e.g. an alkyl iodide) in the presence of an
appropriate base (e.g. an alkali metal alkoxide such as potassium
t-butoxide), advantageously in the presence of a crown ether such as
18-crown-6. Esterification may be effected by reaction with appropriate
acylating agents, such as acyl halides, acid anhydrides and the like.
Useful starting materials for the above compounds of formulae (V), (VII)
and (X) include compounds (XI)
##STR15##
(where R.sup.4 and R.sup.5 are as defined above) and/or 5,6-trans isomers
thereof and the corresponding 1-deoxy compounds; such compounds may be
obtained through oxidative cleavage (e.g. by ozonolysis) of the
22,23-double bond of vitamin D.sub.2, 1.alpha.-hydroxy vitamin D.sub.2 or
O-protected derivatives thereof, these preferably being stabilised by
formation of a Diels Alder dienophile adduct, e.g. with sulphur dioxide or
a diazacyclo compound, for example as described in GB-A-2114570 (the
contents of which are incorporated herein by reference).
Such 20S compounds (XI), optionally still in the form of their dienophile
adducts, may be isomerised by, for example, treatment with a mild base,
e.g. an inorganic base such as sodium bicarbonate or a tertiary organic
base such as 1,4-diazabicyclo ›2.2.2!octane ("DABCO") or 1,8-diazabicyclo
›5.4.0!undec-7-ene ("DBU"). This yields a mixture of 20R and 20S isomers
from which the pure 20R epi-isomer may be isolated chromatographically;
alternatively separation of a desired epi-isomer may be delayed until a
later stage in the synthesis, up to and including the final step.
Reduction of the aldehyde grouping of a compound (XI) or a corresponding
epi-isomer, e.g. using a metal hydride reducing agent such as sodium
borohydride, yields a corresponding hydroxymethyl compound, i.e. a
compound (V) in which Y.sup.a is CH.sub.2 and L is OH. This may be
converted to a compound (V) in which L is a leaving group by, for example,
conversion to a sulphonate ester (e.g. to a tosylate) followed, if
desired, by nucleophilic displacement of the sulphonate group by reaction
with a halide salt (e.g. an alkali metal bromide).
Compounds of formula (V) in which A.dbd. represents a group (A-9) as
hereinbefore defined, Y.sup.a is as hereinbefore defined, and L represents
an O-protected hydroxyl group (e.g. in which the hydroxyl group is
esterified, for example with a lower alkanoyl group such as acetyl) may be
subjected to 1.alpha.-hydroxylation as described under (B) above to give
compounds (V) in which A.dbd. represents a group (A-2) or (A-3) as
hereinbefore defined in which R.sup.5 represents hydrogen. Such compounds
or protected derivatives thereof, e.g. in which R.sup.5 is trimethylsilyl,
may be hydrogenated (e.g. in the presence of a noble metal catalyst such
as tris-triphenylphosphine rhodium chloride) to yield corresponding
compounds in which A.dbd. represents a group (A-4) or (A-5) as
hereinbefore defined, or may be cyclopropanated (e.g. by reaction with
methylene iodide in the presence of zinc/copper couple) to yield
corresponding compounds in which A.dbd. represents a group (A-6) or (A-7)
as hereinbefore defined. Where appropriate, the compounds so obtained may
be converted to compounds in which R.sup.5 is an O-protecting group (e.g.
by silylation) and may be hydrolysed (e.g. with base such as potassium
hydroxide or potassium carbonate) or reduced (e.g. with lithium aluminium
hydride) to remove the side chain ester group to yield useful starting
materials (V) in which L represents a hydroxyl group.
19-Nor analogues of compounds of formula (XI) and corresponding
20-hydroxymethyl compounds (i.e. starting materials for compounds (I) in
which A.dbd. represents a group (A-8) as hereinbefore defined) may be
prepared as described by Perlman et al., Tetrahedron Letters (1992), 33,
pp 2937-2940.
Compounds of formula (V) in which Y.sup.a is e.g. ethylene or trimethylene
may, for example, be obtained by reaction of a compound (V) in which
Y.sup.a is methylene either (i) with a reagent serving to introduce a
one-carbon fragment (e.g. a metal cyanide) and conversion of the group so
introduced to a group --CH.sub.2 L, e.g. by hydrolysing a cyano group to
yield a carboxy group or by reducing such a cyano group (e.g. with a metal
hydride reducing agent such as diisobutyl aluminium hydride) to yield a
carboxaldehyde group, and reducing the carboxy or carboxaldehyde group
(e.g. using sodium borohydride or lithium aluminium hydride) to yield a
hydroxymethyl group which may in turn be subjected to tosylation and, if
desired, nucleophilic displacement as hereinbefore described to effect
conversion to a halomethyl group; or (ii) with a metallated derivative of
an ester or thioester of acetic acid, with a derivative containing another
carbanionic equivalent of acetic acid (e.g. a metallated derivative of
acetonitrile), or with a metallated malonate ester (in which last instance
the reaction product is partially hydrolysed to yield a monoester which
may be decarboxylated by heating to yield a carboxylate ester), reducing
the resulting ester or thioester product to an alcohol (e.g. using lithium
aluminium hydride), and converting the resulting hydroxyl group to a
leaving group, such as a tosylate group or a halogen atom, e.g. as
hereinbefore described.
It will be appreciated that the above procedures (i) and/or (ii) may be
repeated as needed to yield compounds (V) in which Y.sup.a is a C.sub.3
-C.sub.7 alkylene group. D) By reaction of a compound of formula (I) to
modify the substitution pattern about the A.dbd. group, followed if
necessary and/or desired by isomerisation and/or removal of protecting
groups.
Thus for example, compounds (I) in which A.dbd. represents a group (A-4) or
(A-5) may be prepared by hydrogenation of corresponding compounds in which
A.dbd. represents (A-2) or (A-3), e.g. using the method of GB-A-1583749.
It will be appreciated that such hydrogenation may alternatively be
effected at an earlier stage of a reaction sequence, e.g. on a starting
material or intermediate of formula (V).
Compounds (I) in which A.dbd. represents a group (A-6) or (A-7) may be
prepared from corresponding compounds in which A.dbd. represents (A-2) or
(A-3) (in which R.sup.4 is an O-protecting group and R.sup.5 is a hydrogen
atom or a trimethylsilyl group) by Simmons-Smith methylenation (see e.g.
Neef et al., Tetrahedron Letters (1991), 32, pp 5073-5076).
Compounds (I) in which A.dbd. represents a group (A-8) may, for example, be
prepared by cleavage of the 7,8-double bond of an appropriate vitamin D
derivative (e.g. a precursor compound (I) in which A.dbd. is a group
(A-9)), for example by ozonolysis or by successive reaction with potassium
permanganate and sodium periodate, followed by Wittig-Horner reaction of
the resulting 8-one with an appropriate ring A precursor, e.g. of formula
(XII)
##STR16##
(where R.sup.4 and R.sup.5 represent O-protecting groups)--see, for
example, Perlman et al., Tetrahedron Letters (1992), 33, pp 2937-2940.
In general, either 5,6-cis or 5,6-trans geometry may be present at any of
the various steps described in (C) and (D) above, although it may be
preferred to employ 5,6-trans isomers in the above-mentioned
1.alpha.-hydroxylation and 22,23-double bond oxidative cleavage reactions.
Conversion of 5,6-trans geometry to 5,6-cis is thus most advantageously
effected after introduction of the 1.alpha.-hydroxyl group.
It will be appreciated that many of the reaction sequences described above
may also be accomplished using appropriate steroid-5,7-dienes (or
steroid-5-enes which are convertible into such dienes), followed by
conversion of the steroid products into the desired vitamin D analogues,
e.g. by irradiation with UV light.
In general, O-protecting groups present at the 1.alpha.-and/or 3.beta.-
positions may be removed by, for example, conventional methods such as are
well documented in the literature. Thus esterifying acyl groups may be
removed by basic hydrolysis, e.g. using an alkali metal alkoxide in an
alkanol. Etherifying groups such as silyl groups may be removed by acid
hydrolysis or treatment with a fluoride salt, e.g. a tetraalkyl ammonium
fluoride. The use of such acid-labile but base-stable protecting groups
may be of particular advantage during homologation steps to build up a
desired side chain, in view of the strongly basic conditions normally
employed for such reactions.
The following non-limitative examples serve to illustrate the invention.
All temperatures are in .degree.C.
Preparation 1
a)
20.alpha.-Acetoxymethyl-1.alpha.-hydroxy-3.beta.-triisopropylsily-loxy-9,
10-secopregna-5(E),7-diene ›Formula (IV)--A=(A-5). R.sup.3
=.alpha.-CH.sub.3, R.sup.4 =(i-Pr).sub.3 Si , R.sup.5 =H, L=O.CO.CH.sub.3,
Y.sup.a =CH.sub.2 !
A solution of tris-triphenylphosphine rhodium chloride (450 mg) in benzene
(30 ml) (or in a 1:1 mixture of benzene and ethanol) is stirred under
hydrogen until no further uptake is observed. A solution of
20.alpha.-acetoxymethyl-1.alpha.-hydroxy-3.beta.-triisopropylsilyloxy-9,10
-secopregna-5(E), 7,10(19)-triene ›Formula (V)--A=(A-3), R.sup.3
=.alpha.-CH.sub.3, R.sup.4 =(i-Pr).sub.3 Si, R.sup.5 =H, L=O.CO.CH.sub.3,
Y.sup.a =CH.sub.2 --as an alternative the corresponding
1.alpha.-trimethylsilyl ether may be used! (500 mg) in benzene (30 ml) is
added and the mixture stirred under hydrogen until 1 equivalent of
hydrogen has been taken up (ca 21 ml). The title compounds are purified by
chromatography ›the 10(R) and 10(S) isomers may optionally be resolved at
this stage! and have UV A.lambda..sub.max ca. 243,251 and 261 nm, with
.epsilon.=ca. 35,000; 40,000 and 27,000 respectively.
b) 1.alpha.,3.beta.-Bis-triisopropylsilylozy-20.alpha.-hydroxymethyl-9,
10-secopregna-5(E),7-diene ›Formula (V)--A=(A-5), R.sup.3
=.alpha.-CH.sub.3, R.sup.4 =R.sup.5 =(i-Pr).sub.3 Si, L=OH, Y.sup.a
=CH.sub.2 !
The diene from (a) above (ca 500 mg) in dichloromethane (2 ml) is treated
with chlorotriisopropylsilane (250 mg) and imidazole (350 mg) and the
mixture stirred overnight at room temperature. After work up the crude
bis-silyl ether is dissolved in tetrahydrofuran (10 ml), treated with
lithium aluminium hydride (100 mg) and stirred at room temperature for 1-2
hours. After decomposition of the excess lithium aluminium hydride
(careful addition of saturated aqueous sodium sulphate) the reaction
mixture is worked up to afford the title alcohol.
Preparation 2
1.alpha.,3.beta.-Bis-triisoipropylsilyloxy-20.alpha.-hydroxymethyl-9,10-sec
opregna-5(Z),7-diene ›Formula (V)--A=(A-4), R.sup.3 =.alpha.-CH.sub.3,
R.sup.4 =R.sup.5 =(i-Pr).sub.3 Si, L=OH, Y.sup.a =CH.sub.2 !
The 5(E)-triene starting material in Preparation 1(a) is photoisomerised in
benzene in the presence of phenazine by irradiation for 1 hour, to yield
the corresponding 5(Z)-triene. This product is hydrogenated as described
in Preparation 1(a) and silylated and de-acetylated as described in
Preparation 1(b) to give the title compound. UV .lambda..sub.max ca. 243,
251 and 261 nm with .epsilon.=ca. 35,000; 40,000 and 27,000 respectively.
The epi (i.e. 20.beta.-hydroxymethyl) compounds corresponding to the
products of Preparations 1 and 2 are prepared by the same procedures
starting with the 20-epi compound
20.beta.-acetoxymethyl-1.alpha.-hydroxy-3.crclbar.-triisopropylsilyloxy-9,
10-secopregna-5(E),7,10(19)-triene ›Formula (V)--A=(A-3), R.sup.3
=.beta.-CH.sub.3, R.sup.4 =(i-Pr).sub.3 Si, R.sup.5 =H, L=O.CO.CH.sub.3,
Y.sup.a =CH.sub.2 !. This is itself prepared by isomerisation of the
20-aldehyde obtained by ozonolysis of the sulphur dioxide adduct of
vitamin D.sub.2 followed by reduction and 1.alpha.-hydroxylation of the
20-epi aldehyde.
Preparation 3
a)
20.alpha.-Acetoxymethyl-1.alpha.hydroxy-3.beta.-triisonropylsily-loxy-10-s
pirocyclopropyl-9,10-secopregna-5(E),7-diene ›Formula (V)--A=(A-7), R.sup.3
=.alpha.-CH.sub.3, R.sup.4 =(i-Pr).sub.3 Si, R.sup.5 =H, L=O.CO.CH.sub.3,
Y.sup.a =CH.sub.2 !
A mixture of zinc/copper couple (1.08 g) and diiodomethane (0.9 ml) in
ether (6 ml) is heated under reflux with stirring for 40 minutes. A
solution of
20.alpha.-acetoxymethyl-1.alpha.-hydroxy-3.beta.-triisopropylsilyloxy-9,
10-secopregna-5(E), 7,10(19)-triene ›Formula (V)--A=(A-3), R.sup.3
=.alpha.-CH.sub.3, R.sup.4 =(i-Pr).sub.3 Si, R.sup.5 =H, L=O.CO.CH.sub.3,
Y.sup.a =CH.sub.2 --as an alternative the corresponding
1.alpha.-trimethylsilyl ether may be used! (ca. 500 mg) in ether (9 ml) is
added, and the mixture is stirred and heated under reflux until most of
the starting material has disappeared (TLC control: usually about 4 hours
for the 1.alpha.-trimethylsilyl ether, less for the 1.alpha.-hydroxy
compound). The reaction mixture is filtered, the solvent removed and the
product chromatographed to remove the remaining diiodomethane. The title
compound has UV .lambda..sub.max ca. 246, 253 and 263 nm, with
.epsilon.=ca. 29,000; 36,000 and 25,000 respectively.
b)
1.alpha.,3.beta.-Bis-triisopropylsilyloxy-20.alpha.-hydroxymethyl-10-spiro
cyclopropyl-9,10-secopregna-5(E),7-diene ›Formula (V)--A=(A-7), R.sup.3
=.alpha.-CH.sub.3, R.sup.4 =R.sup.5 =(i-Pr).sub.3 Si, L=OH, Y.sup.a
=CH.sub.2 !
The diene from (a) above (ca. 500 mg) in dichloromethane (2 ml) is treated
with chlorotriisopropylsilane (250 mg) and imidazole (350 mg) and the
mixture stirred overnight at room temperature. After work up the crude
bis-silyl ether is dissolved in tetrahydrofuran (10 ml), treated with
lithium aluminium hydride (100 mg) and stirred at room temperature for 1-2
hours. After decomposition of the excess lithium aluminium hydride
(careful addition of saturated aqueous sodium sulphate) the reaction
mixture is worked up to afford the title alcohol.
Preparation 4
1.alpha.,3.beta.-Bis-triisopropylsilyloxy-20.alpha.hydroxymethyl-10-spirocy
clopropyl-9,10-secopregna-5(Z),7-diene ›Formula (V)--A=(A-6) R.sup.3
=.alpha.-CH.sub.3, R.sup.4 =R.sup.5 =(i-Pr).sub.3 Si, L=OH, Y.sup.a
=CH.sub.2 !
The procedure of Preparation 3(a) is repeated starting from the
corresponding 5(Z)-triene, prepared by photoisomerization of the
5(E)-triene as described in Preparation 2; the reaction of the 5(Z)-triene
is somewhat slower than that of the 5(E)-triene. Silylation and
de-acetylation as described in Preparation 3(b) gives the title compound.
UV .lambda..sub.max ca. 246, 253 and 263 nm with .epsilon.=ca. 29,000;
36,000 and 25,000 respectively.
Preparation 5
1.alpha.,3.beta.-Bis-t-butyldimethylsilyloxy-20.beta.-hydroxthyl-19-nor-9,1
0-secopregna-5(E),7-diene ›Formula (V)--A=(A-8), R.sup.3 =.beta.-CH.sub.3,
R.sup.4 =R.sup.5 =t-Bu(Me).sub.2 Si, L=OH, Y.sup.a =CH.sub.2 !
1.alpha.,3.beta.-Bis-t-butyldimethylsilyloxy-20.alpha.-formyl-19-nor-9,10-s
ecopregna-5,7-diene ›Formula (VII)--A=(A-8), R.sup.1 =.alpha.-CH3, R.sup.4
=R.sup.5 =t-Bu(Me).sub.2 Si, Y.sup.b =valence bond! obtained as in
Tetrahedron Lett. (1992), 33, p 2937, (about 1.5g) is dissolved in benzene
(15 ml) and methanol (15 ml) and isomerised by storage overnight with DBU
(400 .mu.l) at 00. The mixture of normal (20.alpha.-formyl) and epi
(20.beta.-formyl) aldehydes may be resolved by chromatography (silica
eluted with 15% benzene in hexane) before or after reduction of the
aldehyde (ca 1 g) in benzene (30 ml) by dropwise treatment with sodium
borohydride, (400 mg) in ethanol (15 ml) at 0.degree., whereafter the
reaction mixture is stirred at 0.degree. for a further 0.5 hour. After
work up the product is resolved by chromatography (silica gel eluting with
benzene or ether in hexane) to yield the title compound.
Preparation 6
a)
1.alpha.,3.beta.-Bis-triisopropylsilyloxy-23-nor-9,10-seco-chola-5(E),7,10
,19-trienic acid, nitrile (mixture of 20-normal and 20- epi isomers)
›Formula (II)--A=(A-3), R.sup.3 =.alpha.- and .beta.-CH.sub.3, R.sup.4
=R.sup.5 =(i-Pr).sub.3 Si, Y.sup.a =CH.sub.2 !
A solution of
1.alpha.,3.beta.-bis-triisopropylsilyloxy-20(.alpha.,.beta.)-tosyloxymethy
l-9,10-secopregna-5(E),7,10(19)-triene ›Formula (V)--A=(A-3), R.sup.3
=.alpha.,.beta.-CH.sub.3, R.sup.4 =R.sup.5 =(i-Pr).sub.3 Si, L=O.tosyl,
Y.sup.a =CH.sub.2 ! (1 g) in dimethylsulphoxide (5 ml) containing
potassium cyanide (390 mg) was heated at 90.degree. for 2 hours, and the
product was extracted (diethyl ether), washed and purified by column
chromatography to give the title nitrile (748 mg). UV (Et.sub.2 O)
.lambda..sub.max 267, .lambda..sub.min 229 nm; NMR (CCl.sub.4)
.delta.5.36-6.13 (ABq, 6,7-H's), 4.83 (bs, 19-H's), 4.13-4.46 (m,
1,3-H's), 0.53 (s, 18-H's).
b)
1.alpha.,3.beta.-Bis-triisopropylsilyloxy-23-nor-9,10-seco-chola-5(E),7,10
,19-trienic carboxaldehyde. (mixture of 20- normal and 20- epi isomers)
›Formula (V)--A=(A-3), R.sup.3 =.alpha.- and .beta.-CH.sub.3, R.sup.4
=R.sup.5 =(i-Pr).sub.3 Si, L=CHO, Y.sup.a =CH.sub.2 !
The nitrile from (a) above (480 mg) in hexane (3 ml) was cooled to
-78.degree. and treated with diisobutylaluminium hydride (1.4 ml of a 1M
solution in heptane). The mixture was stirred at 0.degree. for 1 hour,
treated with ether and saturated ammonium chloride solution, and the
product isolated by extraction into ether. The crude product had UV
(Et.sub.2 O) .lambda..sub.max 270, .lambda..sub.min 229 nm; IR (CCl.sub.4)
.nu..sub.max 1730 cm.sup.-1 ; NMR (CCl.sub.4) .delta. 10.6 (bs, CHO),
5.53-6.23 (ABq, 6,7-H's), 4.76 (bs, 19-H's), 4.16-4.43 (m, 1,3-H's), 56
(s, 18-H's).
c)
1.alpha.,3.beta.-Bis-triisopropylsilyloxy-20(.alpha.,.beta.)-(2-hydroxymet
hyl)-9,10-secopregna-5(E),7,10,19-triene ›Formula (V)--A=(A-3), R.sup.3
=.alpha.- and .beta.-CH.sub.3, R.sup.4 =R.sup.5 =(i-Pr).sub.3 Si, L=OH,
Y.sup.a =(CH.sub.2).sub.2 !
The aldehyde from (b) above (440 mg) in benzenle (10 ml) was treated at
0.degree. with a solution of sodium borohydride (105 mg) in ethanol (10
ml) followed by stirring at room temperature for 45 minutes. After work up
the product was purified by chromatography to give the title compound (380
mg). UV (Et.sub.2 O) .lambda..sub.max 269, .lambda..sub.min 228 nm; IR
(CCl.sub.4) .nu..sub.max 3500-3700 cm.sup.-1 ; NMR (CCl.sub.4) .delta.
5.53-6.3 (ABq, 6,7-H's), 4.73 (bs, 19-H's), 4.16-4.43 (m, 1,3-H's), 0.56
(s, 18-H's).
The isomers (at C-20) were resolved by careful chromatography of 1.2 g of
mixture on silica gel developed with 30% benzene in hexane. The
20.beta.-(epi) isomer (145 mg) was less polar and eluted first followed by
a mixture of isomers and then the 20.alpha.-(normal) isomer (360 mg).
d)
1.alpha.,3.beta.-Bis-triisoropylsilyloxy-20.alpha.-(2-bromoethyl)-9,10-sec
opregna-5(E),7,10(19)-triene ›Formula (V)--A=(A-3), R.sup.3
=.alpha.-CH.sub.3, R.sup.4 =R.sup.5 =(i-Pr).sub.3 Si, L=Br, Y.sup.a
=(CH.sub.2).sub.2 !
The normal alcohol from (c) above (200 mg) was stirred at room temperature
for 2 hours in dichloromethane (5 ml) containing p-toluenesulphonyl
chloride (110 mg) and pyridine (243 .mu.l). Sodium bicarbonate (20 ml of a
saturated solution) was added, the stirring continued for a further 2
hours, and the reaction mixture worked up. The crude tosylate was
dissolved in acetonitrile (6.6 ml) and dichloromethane (6.6 ml) containing
lithium bromide (317 mg) and 1,8 bis-dimethylaminonaphthalene ("proton
sponge" 40 mg) and the mixture heated under reflux at 80.degree. for 30
minutes. The mixture was then cooled and worked up to give the title
bromide (261 mg, purified by chromatography). UV (Et.sub.2 O)
.lambda..sub.max 267, .lambda..sub.min 228 nm; NMR (CCl.sub.4) .delta.
5.43-6.16 (ABq, 6,7-H's), 4.76 (bs, 19-H's), 4.14-4.45 (m, 1,3-H's), 3.16
(m, Cl.sub.2 Br), 0.5 (s, 18-H's).
Preparation 7
a)
1.alpha.,3.beta.-Bis-triisopropylsilyloxy-20.alpha.-bromomethyl-9,10-secop
regna-5(E),7-diene ›Formula (V)--A=(A-5), R.sup.3 =.alpha.-CH.sub.3,
R.sup.4 =R.sup.5 =(i-Pr).sub.3 Si, L=Br, Y.sup.a =CH.sub.2 !
This compound is prepared from the product of Preparation 1 following the
procedure of Preparation 6(d).
b)
1.alpha.,3.beta.-Bis-triisopropylsilyloxy-20.alpha.-bromomethy-9,10-secopr
egna-5(Z),7-diene ›Formula (V)--A=(A-4), R.sup.3 =.alpha.-CH.sub.3, R.sup.4
=R.sup.5 =(i-Pr).sub.3 Si, L=Br, Y.sup.a =CH.sub.2 !
This compound is prepared from the product of Preparation 2 following the
procedure of Preparation 6(d).
c)
1.alpha.,3.beta.-Bis-triisopropylsilyloxy-20.alpha.-bromoethyl-10-spirocyc
lopropyl-9,10-secopregna-5(E),7-diene ›Formula (V)--A=(A-7), R.sup.3
=.alpha.-CH.sub.3, R.sup.4 =R.sup.5 =(i-Pr).sub.3 Si, L=Br, Y.sup.a
=CH.sub.2 !
This compound is prepared from the product of Preparation 3 following the
procedure of Preparation 6(d).
d)
1.alpha.,3.beta.-Bis-triisopropylsilyloxy-20.alpha.-bromoethyl-10-spirocyc
lopropyl-9,10-secopregna-5(Z),7-diene ›Formula (V)--A=(A-6), R.sup.3
=.alpha.-CH.sub.3, R.sup.4 =R.sup.5 =(i-Pr).sub.3 Si, L=Br, Y.sup.a
=CH.sub.2 !
This compound is prepared from the product of Preparation 4 following the
procedure of Preparation 6(d).
e)
1.alpha.,3.beta.-Bis-t-butyldimethylsilyloxy-20.beta.-bromomethyl-12-nor-9
,10-secopregna-5(E),7-diene ›Formula (V)--A=(A-8), R.sup.3
=.beta.-CH.sub.3, R.sup.4 =R.sup.5 =(i-Pr).sub.3 Si, L=Br. Y.sup.a
=CH.sub.2 !
This compound is prepared from the product of Preparation 5 following the
procedure of Preparation 6(d).
Preparation 8
a)
1.alpha.,3.beta.-Bis-triisopropylsilyloxy-20.alpha.-bromoethyl-9,10-secopr
egna-5(E),7-diene ›Formula (V)--A=(A-5), R.sup.3 =.alpha.-CH.sub.3, R.sup.4
=R.sup.5 =(i-Pr).sub.3 Si, L=Br, Y.sup.a =(CH.sub.2).sub.2 !
The title compound is prepared from the product of Preparation 7(a)
following the procedures of Preparation 6(a)-(d).
b)
1.alpha.,3.beta.-Bis-triisopropylsilyloxy-20.alpha.-bromoethyl-9,10-secopr
egna-5(Z),7-diene ›Formula (V)--A=(A-4), R.sup.3 =.alpha.-CH.sub.3, R.sup.4
=R.sup.5 =(i-Pr)-.sub.3 Si, L=Br, Y.sup.a =(CH.sub.2).sub.2 !
The title compound is prepared from the product of Preparation 7(b)
following the procedures of Preparation 6(a)-(d).
c)
1.alpha.,3.beta.-Bis-triisopropylsilyloxy-20.alpha.-bromoethyl-10-spirocyc
lopropyl-9,10-secopreana-5(E),7-diene ›Formula (V) A=(A-7), R.sup.3
=.alpha.-CH.sub.3, R.sup.4 =R.sup.5 =(i-Pr).sub.3 Si, L=Br, Y.sup.a
=(CH.sub.2).sub.2 !
The title compound is prepared from the product of Preparation 7(c)
following the procedures of Preparation 6(a)-(d).
d)
1.alpha.,3.beta.-Bis-triisopropylsilyloxy-20.alpha.-bromoethyl-10-spirocyc
lopropyl-9,10-secopregna-5(Z),7-diene ›Formula (V)--A=(A-6). R.sup.3
=.alpha.-CH.sub.3, R.sup.4 =R.sup.5 =(i-Pr).sub.3 Si, L=Br, Y.sup.a
=(CH.sub.2).sub.2 !
The title compound is prepared from the product of Preparation 7(d)
following the procedures of Preparation 6(a)-(d).
e)
1.alpha.,3.beta.-Bis-t-butyldimethylsilyloxy-20.beta.-bromoethyl-19-nor-9,
10-secopregna-5,7-diene ›Formula (V)--A=(A-8), R.sup.3 =.beta.-CH.sub.3,
R.sup.4 =R.sup.5 =(i-Pr).sub.3 Si, L=Br, Y.sup.a =(CH.sub.2).sub.2 !
The title compound is prepared from the product of Preparation 7(e)
following the procedures of Preparation 6(a)-(d).
Preparation 9
a)
1.alpha.,3.beta.-Bis-triisopropylsilyloxy-24-carbamoyl-24-homo-9,10-secoch
ola-5(E),7,10(19),22(E), 24(E)-pentaene ›NH.sub.2 amide of acid of formula
(X)--A=(A-3), R.sup.3 =.alpha.-CH.sub.3, R.sup.4 =R.sup.5 =(i-Pr).sub.3
Si, Y=CH.dbd.CH--CH.dbd.CH!
Lithium aluminium hydride (10 ml of a 1M solution in ether) was stirred at
room temperature under an atmosphere of ammonia for 30 minutes. The
resulting suspension was treated with
1.alpha.,3.beta.-bis-triisopropylsilyloxy-24-ethoxycarbonyl-24-homo-9,10-s
ecochola-5(E), 7,10(19),22(E),24(E)-pentaene ›ethyl ester of acid of
formula (X)--A=(A-3), R.sup.3 =.alpha.-CH.sub.3, R.sup.4 =R.sup.5
=(i-Pr).sub.3 Si, Y=CH.dbd.CH--CH.dbd.CH! (250 mg) in ether (1 ml) and the
mixture was stirred for 10 hours at room temperature. The reaction mixture
was then cooled to 0.degree., treated (caution|) with aqueous ethanol (3
ml, 70%), diluted with ether and dried over sodium sulphate. Purification
by column chromatography gave the title compound (180 mg). UV (Et.sub.2 O)
.lambda..sub.max 260 nm; IR .nu..sub.max (CDCl.sub.3) 3520-3000, 1670,
1630, 1580 cm.sup.-1 ; NMR (CDCl.sub.3) .delta. 0.63 (s, 18-H's), 3.8-4.7
(m, 1,3-H's), 4.7-5.0 (bs, 19-H's), 5.3-7.3 (m, 6,7,22,23,24,24a-H's).
b)
1.alpha.,3.beta.-Bis-triisopropylsilyloxy-24-cyano-24-homo-9,10-secochola-
5(E),7,10(19),22(E),24(E)-pentaene ›Formula (II)--A=(A-3), R.sup.3
=.alpha.-CH.sub.3, R.sup.4 =R.sup.5 =(i-Pr).sub.3 Si,
Y=CH.dbd.CH--CH.dbd.CH!
Trifluoroacetic anhydride (54 .mu.l) was added dropwise to a stirred,
ice-cooled solution of the amide from (a) above in anhydrous dioxan (540
.mu.l) containing pyridine (150 .mu.l). The mixture was stirred for 1 hour
at room temperature, diluted with ether, washed successively with water,
aqueous hydrochloric acid, saturated aqueous sodium bicarbonate and brine
and then dried. The solvents were removed in vacuo and the product was
isolated by chromatography to give the title compound (115 mg). UV
(Et.sub.2 O) .lambda..sub.max 260 nm; IR .nu..sub.max (CDCl.sub.4) 2100,
1630 cm.sup.-1 ; NMR (CDCl.sub.3) .delta. 0.56 (s, 18-H's), 3.8-4.6 (m,
1,3-H's), 4.6-5.0 (bs, 19-H's), 5.3-7.3 (m, 6,7,22,23, 24,24a-H's).
EXAMPLE 1
a)
1.alpha.,3.beta.-Bis-triisopropylsilyloxy-9,10-secochola-5(E),7,10(19)-tri
ene-24-carbonitrile ›Formula (II)--A=(A-3), R.sup.3 =.alpha.-CH.sub.3,
R.sup.4 =R.sup.5 =(i-Pr).sub.3 Sii, Y=(CH.sub.2).sub.3 !
A solution of
1.alpha.,3.beta.-bis-triisopropylsilyloxy-24-tosyloxy-9,10-secochola-5(E),
7,10(19)-triene, generated in situ by tosylation of the corresponding 24-ol
(480 mg, prepared as described in Example 2(c) of WO 93/09093), in
dimethyl sulphoxide (2.5 ml) containing potassium cyanide (220 mg) was
heated at 90.degree. for 50 minutes, cooled and extracted with ethyl
acetate. The thus-obtained product was purified by column chromatography
to yield the title compound (320 mg). UV (Et.sub.2 O) .lambda..sub.max
267, .lambda..sub.min 236 nm; NMR (CCl.sub.4) .delta. 5.53-6.3 (ABq,
6,7-H's), 4.8 (s, 19-H's), 0.50 (s, 18-H's).
b)
25-Amino-1.alpha.,30-bis-triisopropylsilyloxy-9,10-secocholesta-5(E),7,10(
19)-triene ›Formula (I)--A=(A-3), R=H, R.sup.1 =R.sup.2 =CH.sub.3, R.sup.3
=.alpha.-CH.sub.3, R.sup.4 =R.sup.5 =(i-Pr).sub.3 Sii, Y=(CH.sub.2).sub.3
!
Tetrahydrofuran (2 ml) was added to cerous chloride (639 mg) at 0.degree..
The resulting solution was stirred at room temperature for 15 minutes,
cooled to -70.degree., treated with methyl lithium (1.85 ml of a 1.6M
solution in tetrahydrofuran) and stirred briefly at -70.degree.. The
nitrile from (a) above (320 mg) in tetrahydrofuran (2.5 ml) was added at
-70.degree. to the thus-prepared alkyl cerium reagent and the reaction
mixture was stirred for 1 hour at -70.degree., briefly warmed to room
temperature, cooled to -700.degree., quenched with concentrated aqueous
ammonia (2 ml) and filtered through Celite, which was thereafter washed
with tetrahydrofuran and diethyl ether. The product was isolated from the
combined organic phases and purified by chromatography (alumina) to give
the title compound (235 mg). UV (Et.sub.2 O) .lambda..sub.max 268,
.lambda..sub.min 228 nm; NMR (CDCl.sub.3) .delta. 5.58-6.3 (ABq, 6,7-H's),
4.83 (s, 19-H's), 0.53 (s, 18-H's).
c)
25-Amino-1.alpha.,3.beta.-bis-triisopropylsilyloxy-9,10-secocholesta-5(Z),
7,10(19)-triene ›Formula (I)--A=(A-2). R=H, R.sup.1 =R.sup.2 =CH.sub.3
R.sup.3 =-CH.sub.3, R.sup.4 =R.sup.5 =(i-Pr).sub.3 SiSi,
Y=(CH.sub.2).sub.3 !
A solution of the 5(E) compound from (b) above (188 mg) in benzene (25 ml)
containing phenazine (98 mg) was irradiated for 50 minutes. The product
was worked up and purified by TLC to give the title compound (140 mg). UV
(Et.sub.2 O) .lambda..sub.max 260, .lambda..sub.min 222 nm; NMR
(CDCl.sub.3) 6 5.66-6.2 (ABq, 6,7-H's), 4.7-5.03 (d, 19-H's), 0.50 (s,
18-H's).
d)
25-Amino-1.alpha.,3.beta.-dihydroxy-9,10-secocholesta-5(Z),7,10(19)-triene
›Formula (I)--A=(A-2), R=H, R.sup.1 =R.sup.2 =CH.sub.3 R.sup.3 =-CH.sub.3,
R.sup.4 =R.sup.5 =H, Y=(CH.sub.2).sub.3 !
The bis-silyl ether from (c) above (60 mg) in tetrahydrofuran (0.556 .mu.l)
was desilylated by treatment with tetrabutylammonium fluoride (0.556 .mu.l
of a 1M solution in tetrahydrofuran) for 3 hours. The product was
extracted into chloroform, which was then washed twice with water, dried
and evaporated in vacuo. The thus-obtained product was purified by two
successive TLC's to yield the title compound (10.6 mg). UV (EtOH)
.lambda..sub.max 264, .lambda..sub.min 228nm; NMR (CDCl.sub.3) .delta.
5.66-6.23 (ABq, 6,7-H's), 4.8-5.13 (d, 19-H's), 1.1-1.2 (d, 26,27-H's),
0.50 (s, 18-H's) ; IR (CDCl.sub.3) .nu..sub.max 3600-3350 cm.sup.-1
(OH,NH)
EXAMPLE 2
a)
1.alpha.,3.beta.-Bis-triisopropylsilyloxy-22,23-bisnor-9,10-secochola-5(E)
,7,10(19)-triene-24-carbonitrile, mixture of 20R and 20S isomers ›Formula
(II)--A=(A-3), R.sup.3 =.alpha.-and .beta.-CH.sub.3 (.about.1:1), R.sup.4
=R.sup.5 =(i-Pr).sub.3 Sii, Y=CH.sub.2 !
A solution of
1.alpha.,3D-bis-triisopropylsilyloxy-20(.alpha.,.beta.)-tosyloxymethyl-9,1
0-secopregna-5(E),7,10(19)-triene ›Formula (V)--A=(A-3), R.sup.3 =.alpha.-
and .beta.-CH.sub.3, R.sup.4 =R.sup.5 =(i-Pr).sub.3 Sii, Y.sup.a
=CH.sub.2, L=tosyloxy! (1 g) in dimethylsulphoxide (5 ml) containing
potassium cyanide (390 mg) was heated at 90.degree. for 2 hours. The
product was extracted with diethyl ether, washed and purified by column
chromatography to give the title compound (748 mg). UV (Et.sub.2 O)
.lambda..sub.max 267, .lambda..sub.min 229 nm; NMR (CCl.sub.4) .delta.
5.36-6.13 (ABq, 6,7-H's), 4.83 (bs, 19-H's), 4.13-4.46 (m, 1,3-H's), 0.53
(s, 18-H's).
b)
25-Amino-1.alpha.,3.beta.-bis-triisopropylsilyloxy-23.24-bisnor-9,10-secoc
holesta-5(E),7,10(19)-triene, mixture of 20R and 20S isomers ›Formula
(I)--A=(A-3), R=H, R.sup.1 =R.sup.2 =CH.sub.3, R.sup.3 =.alpha.- and
.beta.-CH.sub.3 (.about.1.1), R.sup.4 =R.sup.5 =(i-Pr).sub.3 Sii,
Y=CH.sub.2 !
A solution of cerous chloride (492 mg, 2 mM) in tetrahydrofuran (2.5 ml)
was stirred at room temperature for 1 hour, cooled to -78.degree., treated
with methyl lithium (2 mM in hexane), and stirred for a further 30 minutes
at -78.degree.. A solution of the product from (a) above (262 mg) in
tetrahydrofuran (1.5 ml) was added, and the reaction mixture was stirred
at -78.degree. for 1.5 hours, allowed to warm to room temperature over 2
hours, cooled again to -78.degree., and worked up as in Example 1(b) to
give the title compound (170 mg). UV (Et.sub.2 O) .lambda..sub.max 269 nm;
NMR (CDCl.sub.3) .delta. 5.53-6.33 (ABq, 6,7-H's), 4.76 (s, 19-H's), 0.53
(s, 18-H's).
c)
25-Amino-1.alpha.,3.beta.-bis-triisopropylsilyloxy-23.24-bisnor-9,10-secoc
holesta-5(Z),7,10(19)-triene, mixture of 20R and 20S isomers ›Formula
(I)--A=(A-2), R=H, R.sup.1 =R.sup.2 =CH.sub.3, R.sup.3 =.alpha.- and
.beta.-CH.sub.3 (.about.1:1), R.sup.4 =R.sup.5 =(i-Pr).sub.3 Sii,
Y=CH.sub.2 !
A solution of the product from (b) above (170 mg) in benzene (20 ml)
containing phenazine (80 mg) was photoisomerised and worked up as in
Example 1(c) to give the title compound (90 mg). UV (Et.sub.2 O)
.lambda..sub.max 262 nm; NMR (CDCl.sub.3) .delta. 5.63-6.06 (ABq,
6,7-H's), 4.9-5.2 (each s, 19-H's), 1.4 (s, gem CH.sub.3 's), 0.50 (s,
18-H's).
d)
25-Amino-1.alpha.,3.beta.-dihydroxy-23.24-bisnor-9,10-secocholesta-5(Z),7,
10,(19)-triene, mixture of 20R and 20S isomers ›Formula (I)--A=(A-2), R=H,
R.sup.1 =R.sup.2 =CH.sub.3, R.sup.3 =.alpha.-and .beta.-CH.sub.3
(.about.1:1), R.sup.4 =R.sup.5 =H, Y=CH.sub.2 !
A solution of the product from (c) above (90 mg) in tetrahydrofuran (0.5
ml) was stirred with tetrabutylammonium fluoride (0.38 ml of a 1M solution
in tetrahydrofuran) overnight. Monitoring by TLC showed that unchanged
starting material remained, so the mixture was treated with further
tetrabutylammonium fluoride (0.65 ml) and stirred for a further 3 hours.
The mixture was worked up and the product was isolated and purified by
column chromatography on alumina (twice) to give the title compound. UV
(EtOH) .lambda..sub.max 263 nm; IR (CDCl.sub.3) .nu..sub.max 3600-3340
cm.sup.-1 (OH, NH); NMR (CDCl.sub.3) .delta. 5.56-6.23 (ABq, 6,7-H's),
4.9-5.23 (each s, 19-H's), 1.03, 1.23, 1.36 (m, 21-CH.sub.3, gem CH.sub.3
's), 0.56 (s, 18-H's)
EXAMPLE 3
a)
25-Acetamido-1.alpha.,3.beta.-bis-triisopropylsilyloxy-9,10-secocholesta-5
(Z),7,10(19)-triene ›Formula (I)--A=(A-2), R=CH.sub.3 CO, R.sup.1 =R.sup.2
=CH.sub.3, R.sup.3 =.alpha.-CH.sub.3, R.sup.4 =R.sup.5 =(i-Pr).sub.3 Sii,
Y=(CH.sub.2).sub.3 !
A solution of the product from Example 1(c) (60 mg) in methylene chloride
(1 ml) was treated with acetic anhydride (290 .mu.l) and pyridine (290
.mu.l) and the resulting mixture was stirred at room temperature for 3.5
hours then treated with aqueous sodium bicarbonate. After a further 2
hours the mixture was worked up and the product was isolated by TLC to
give the title compound (42 mg). UV (Et.sub.2 O) .lambda..sub.max 261,
.lambda..sub.min 225 nm; IR (CCl.sub.4) 3420, 3300 (NH), 1670 (C=0)
cm.sup.-1 ; NMR (CCl.sub.4) .delta. 5.96-6.1 (ABq, 6,7-H's), 4.76, 5.06
(each s, 19-H's), 1.76 (s, COCH.sub.3), 1.23 (s, gem CH.sub.3 's), 0.58
(s, 18-H's).
b)
25-Acetamido-1.alpha.,3.beta.-dihydroxy-9,10-secocholesta-5(Z),7,10(19)-tr
iene ›Formula (I)--A=(A-2), R=CH.sub.3 CO R.sup.1 =R.sup.2 =CH.sub.3,
R.sup.3 =.alpha.-CH.sub.3, R.sup.4 =R.sup.5 =H, Y=(CH.sub.2).sub.3 !
A solution of the bis-silyl ether from (a) above (42 mg) in tetrahydrofuran
(0.37 ml) was treated with tetrabutylammonium fluoride (0.36 ml of a 1M
solution in tetrahydrofuran). The resulting mixture was stirred at room
temperature for 2 hours, treated with further tetrabutylammonium fluoride
solution (0.1 ml), stirred for 3 hours and worked up. The product was
isolated by TLC to give the title compound (20.2 mg). UV (EtOH)
.lambda..sub.max 263, .lambda..sub.min 225 nm; IR (CDCl.sub.3) 3420, 3600
(NH, OH), 1660 (C=0) cm.sup.-1 ; NMR (CDCl.sub.3) .delta. 5.7-6.3 (ABq,
6,7-H's), 4.8, 5.06 (each s, 19-H's), 1.83 (s, COCH.sub.3), 1.23 (s, gem
CH3's), 0.83, 0.9 (d, 21-H's), 0.58 (s, 18-H's).
EXAMPLE 4
a)
1.alpha.,3.beta.-Bis-triisopropylsilyloxy-20-epi-9,10-secochola-5(E),7,10(
19)-triene-24-carbonitrile ›Formula (II) A=(A-3-, R.sup.3 =.beta.-CH.sub.3,
R.sup.4 =R.sup.5 =(i-Pr).sub.3 Si, Y=(CH.sub.2).sub.3 !
Acetonitrile (0.32 ml) in tetrahydrofuran (2 ml) was added dropwise at
-78.degree. to a solution of butyl, lithium (3.75 ml) of a 1.6M solution
in hexane) and tetrahydrofuran (4 ml). After 50 minutes storage at
78.degree. all but a 0.38 mMole portion of the solution (presumed to
contain a total of 6 mMoles) was expelled and the remaining portion
treated with a solution of
1.alpha.,3.beta.-bis-triisopropylsilyoxy-20.beta.-bromoethyl-9,10-secopreg
na-5(E), 7,10(19)-triene ›Formula (V)--A=(A-3),R.sup.3 =.beta.-CH.sub.3
R.sup.4 =R.sup.5 =(i-Pr).sub.3 Si, L=Br, Y=(CH.sub.2).sub.2 ! (100 mg) in
tetrahydrofuran (2.05 ml). After 1.5 hours storage at -780.degree.
(starting material no longer present by TLC), the reaction mixture was
treated with aqueous ammonium chloride and the product extracted into
ether. The crude product was combined with similar material from a second
reaction (carried out as above on 172 mg bromide) and the product purified
by chromatography to give the title compound (193 mg). (UV (Et.sub.2 O)
.lambda..sub.max 269, .lambda..sub.min 228 nm; NMR (CCl.sub.4) .delta.
0.53 (s,18-H's), 4.8 (s, 19-H's), 5.56-6.3 (ABq, 6,7-H's).
b)
25-Amino-1.alpha.,3.beta.-bis-triisopropylsilylozy-20-epi-9,10-secocholest
a-5(E),7,10(19)-triene ›Formula (I)--A=(A-3), R=H, R.sup.1 =R.sup.2
=CH.sub.3, R.sup.4 =R.sup.5 =(i-Pr).sub.3 Si, Y=(CH.sub.2).sub.3 !
A solution of cerous chloride in tetrahydrofuran was treated at 0.degree.
with sufficient methyl lithium (1.8 ml of a ›?M! solution in hexane) to
produce a persistent yellow colour. The solution was cooled to
-78.degree., a further portion of methyl lithium (1.35 ml) was added and
the mixture was kept at -78.degree. for 40 minutes to complete formation
of the organocerium reagent. The nitrile from (a) above (196 mg in 3 ml
tetrahydrofuran) was added at -78.degree. and the reaction mixture was
stirred at that temperature for an additional 60 minutes, warmed to
-40.degree., cooled to -78.degree. and treated with ammonium hydroxide.
The crude product was filtered through celite (methylene chloride/diethyl
ether) and purified by chromatography to give the title compound (120 mg).
UV (Et.sub.2 O), .lambda..sub.max 269, .lambda..sub.min 228 nm; NMR
(CCl.sub.4) .delta. 0.53 (s, 18-H's), 4.13-4.66 (bm,1,3-H's), 4.9 (s,
19-H's), 5.66-6.46 (ABq,6,7-H's), 6.46 (bs, NH's, exchanges with D.sub.2
O).
c)
25-Amino-1.alpha.,3.beta.-bis-triisopropylsilyloxy-20-epi-9-secocholesta-5
(Z),7,10(19)-triene ›Formula (I)--A=(A-2), R=H, R.sup.1 =R.sup.2 =CH.sub.3,
R.sup.3 =.beta.-CH.sub.3 R.sup.4 =R.sup.5 =(i-Pr).sub.3 Si,
Y=(CH.sub.2).sub.3 !
The amine from (b) above (80 mg) was photoisomerised by irradiation for 40
minutes in solution in benzene (10.6 ml) contained phenazine (42 mg).
Chromotography afforded the title compound (50 mg). UV (Et.sub.2 O)
.lambda..sub.max 263, .lambda..sub.min 226 nm; NMR (CDCl.sub.3) .delta.
0.50 (s, 18-H's), 4.06-4.6 (bm, 1,3-H's), 4.6. 4.76 (ea. s, 19-H's),
5.53-6.2 (ABq, 6,7-H's), 6.2 (bs, NH's, exchanges with D.sub.2 O).
d)
25-Amino-1.alpha.,3.beta.-dihydroxy-20-epi-9,10-secocholesta-5(Z),7,10(19)
-triene ›Formula (I)--A=(A-2), R=H, R.sup.1 =R.sup.2 =CH.sub.3, R.sup.3
=.beta.-CH.sub.3, R.sup.4 =R.sup.5 =H, Y=(CH.sub.2).sub.3 !
The silyl ether (52 mg) from (c) above was desilyated by treatment with
tetrabutylammonium fluoride (350 .mu.l) in tetrahydrofuran (350 .mu.l)
overnight at room temperature. Chromatography gave the title compound (8.7
mg). UV (EtOH) .lambda..sub.max 263-4, .lambda..sub.min 225-6 nm; IR
(CHCl.sub.3) 3400, 3600cm-.sup.1 (OH, NH); NMR (CDCl.sub.3) .delta. 0.53
(s, 18-H's), 0.86, 0.76 (d 21-H's), 2.03 (bs, NH's, exchanges with D.sub.2
O), 4.06-4.46 (bm, 1,3-H's), 4.9, 5.23 (ea. s, 19-H's), 5.76-6.4 (ABq,
6,7-H's).
The compound
25-amino-1.alpha.,3.beta.-dihydroxy-9,10-secocholesta-5(Z),7,10(19)-triene
›Formula (I)--A =(A-2), R=H, R.sup.1 =R.sup.2 =CH.sub.3, R.sup.3
=.alpha.-CH.sub.3, R.sup.4 =R.sup.5 =H, Y=(CH.sub.2).sub.3 ! is prepared
by similar reaction according to steps (a)-(d) above of the product of
Preparation 6(d).
The compound
25-amino-1.alpha.,3.beta.-dihydroxy-9,10-secocholesta-5(E),7-diene
›Formula (I)--A=(A-5), R=H, R.sup.1 =R.sup.2 =CH.sub.3, R.sup.3
=.alpha.-CH.sub.3, R.sup.4 =R.sup.5 =H, Y=(CH.sub.2).sub.3 ! is prepared
by similar reaction according to steps (a)-(d) above of the product of
Preparation 8(a).
The compound
25-amino-1.alpha.,3.beta.-dihydroxy-9,10-secocholesta-5(Z),7-diene
›Formula (I)--A=(A-4), R=H, R.sup.1 =R.sup.2 =CH.sub.3, R.sup.3
=.alpha.-CH.sub.3, R.sup.4 =R.sup.5 =H, Y=(CH.sub.2).sub.3 ! is prepared
by similar reaction according to steps (a)-(d) above of the product of
Preparation 8(b).
The compound
25-amino-1.alpha.,3.beta.-dihydroxy-10-spirocyclopropyl-9,10-secocholesta-
5(E),7-diene ›Formula (I)--A=(A-7), R=H, R.sup.1 =R.sup.2 =CH.sub.3,
R.sup.3 =.alpha.-CH3, R.sup.4 =R.sup.5 =H, Y=(CH.sub.2).sub.3 ! is
prepared by similar reaction according to steps (a)-(d) above of the
product of Preparation 8(c).
The compound
25-amino-1.alpha.,3.beta.-dihydroxy-10-spirocyclopropyl-9,10-secocholesta-
5(Z),7-diene (Formula (I)--A=(A-6), R=H, R.sup.1 =R.sup.2 =CH.sub.3,
R.sup.3 =.alpha.-CH.sub.3, R.sup.4 =R.sup.5 =H, Y=(CH.sub.2).sub.3 ! is
prepared by similar reaction according to steps (a)-(d) above of the
product of Preparation 8(d).
The compound
25-amino-1.alpha.,3.beta.-dihydroxy-20-epi-19-nor-9,10-secocholesta-5,7-di
ene ›Formula (I)--A=(A-8), R=H, R.sup.1 =R.sup.2 =CH.sub.3, R.sup.3
=.beta.-CH.sub.3, R.sup.4 =R.sup.5 =H, Y=(CH.sub.2).sub.3 ! is prepared by
similar reaction according to steps (a)-(d) above of the product of
Preparation 8(e).
EXAMPLE 5
a)
1.alpha.,3.beta.-Bis-triisopropylsilyloxy-24-homo-9,10-secochola-5(E),7,10
(19)-triene-24-carbonitrile ›Formula (II)--A=(A-3), R.sup.3
=.alpha.-CH.sub.3 R.sup.4 =R.sup.5 =(i-Pr).sup.3 Si, Y=(CH.sub.2).sub.4 !
Acetonitrile (0.32 ml) in tetrahydrofuran (2 ml) was added dropwise at
-78.degree. to a solution of butyl lithium (3.75 ml of a 1.6M solution in
hexane) and tetrahydrofuran (4 ml). After 50 minutes storage at
-78.degree. all but a 0.7 mMole portion of the solution (presumed to
contain a total of 6 mMoles) was expelled and the remaining portion was
treated with a solution of
1.alpha.,3-bis-triisopropylsilyloxy-24-bromo-9,10-secochola-5(E),7,10(19)-
triene ›Formula (V)--A=(A-3), R.sup.3 =.alpha.-CH.sub.3, R.sup.4 =R.sup.5
=(i-Pr).sub.3 Si, L=Br, Y=(CH.sub.2).sub.3 ! (190 mg) in tetrahydrofuran
(3.05 ml). After 40 minutes storage at -780.degree. the mixture was
allowed to warm to -30.degree., kept at that temperature for 60 minutes
(starting material no longer present by TLC), then cooled to -78.degree..
Thereafter the reaction mixture was treated with aqueous ammonium chloride
and the product was extracted into diethyl ether. The crude product was
purified by chromatography to give the title compound (156 mg). UV (EtOH)
.lambda..sub.max 267-8, .lambda..sub.min 228 nm; NMR (CCl.sub.4) .delta.
0.53 (s, 18-H's), 4.96, (s, 19-H's), 5.53-6.26 (ABq, 6,7-H's).
b)
25-Amino-1.alpha.,3.beta.-bis-triisopropylsilyloxy-24-homo-9,10-secocholes
ta-5(E),7,10(19)-triene ›Formula (I)--A=(A-3), R=H, R.sup.1 =R.sup.2
=CH.sub.3, R.sup.3 =.alpha.-CH.sub.3 R.sup.4 =R.sup.5 =(i-Pr).sub.3 Si,
Y=(CH.sub.2).sub.4 !
A solution of cerous chloride (381 mg) in tetrahydrofuran (3 ml) was
treated at 0.degree. with sufficient methyl lithium (1.2 mL of a ›?! molar
solution in hexane) to produce a persistent yellow colour. The solution
was cooled to -78.degree., a further portion of methyl lithium (1.4 ml)
was added and the mixture was kept at -78.degree. for 30 minutes to
complete formation of the organocerium reagent. The nitrile from (a) above
(180 mg in 2.05 ml tetrahydrofuran) was added at -78.degree., and the
reaction mixture was stirred at that temperature for an additional 60
minutes, warmed to -30.degree., cooled to -78.degree., and treated with
aqueous ammonium hydroxide. The crude product was filtered through Celite
(methylene chloride/ diethyl ether) and purified by chromatography to give
the title compound (125 mg). UV (Et.sub.2 O) .lambda..sub.max 267-8,
.lambda..sub.min 227-8 nm; NMR (CCl.sub.4) .delta. 0.53 (s, 18-H's), 4.8,
(s, 19-H's), 5.53-6.26 (ABq, 6,7-H's).
c)
25-Amino-1.alpha.,3.beta.-bis-triisopropylsilyloxy-24-homo-9,10-secocholes
ta-5(Z),7,10(19)-triene ›Formula (I)--A=(A-2), R=H, R.sup.1 =R.sup.2
=CH.sub.3,R.sup.3 =.alpha.-CH.sub.3 R.sup.4 =R.sup.5 =(i-Pr).sub.3 Si,
Y=(CH.sub.2).sub.4 !
The amine from (b) above (125 mg) was photoisomerised by irradiation for 40
minutes in solution in benzene (18 ml) containing phenazine (61 mg).
Chromatography afforded the title compound (88 mg). UV (Et.sub.2 O)
.lambda..sub.max 263, .lambda..sub.min 226-7 nm; NMR (CDCl.sub.3) .delta.
0.53 (s, 18-H's), 4.73, 5.0 (ea. s, 19-H's), 5.6-6.1 (ABq, 6,7-H's).
d)
25-Amino-1.alpha.,3.beta.-dihydroxy-24-homo-9,10-secocholesta-5(Z),7,10(19
)-triene ›Formula (I)--A=(A-2), R=H, R.sup.1 =R.sup.2 =CH.sub.3, R.sup.3
=.alpha.-CH.sub.3, R.sup.4 =R.sup.5 =H, Y=(CH.sub.2).sub.4 !
The silyl ether (40 mg) from (c) above was desilyated by treatment with
tetrabutylammonium fluoride (320 .mu.l) in tetrahydrofuran (320 .mu.l) for
three hours at room temperature. Chromatography gave the title compound
(8.7 mg). UV (EtOH) .lambda..sub.max 263, .lambda..sub.min 226-7 nm; IR
(CHCl.sub.3) .nu..sub.max 3200-3300, 3600 cm.sup.-1 (OH, NH); NMR
(CDCl.sub.3) .delta. 0.53 (s, 18-H's), 4.9, 5.2 (ea. s, 19-H's), 5.8-6.33
(ABq, 6,7-H's).
e)
25-Amino-1.alpha.,3.beta.-dihydroxy-24,26,27-tris-homo-9,10-secocholesta-5
(Z),7,10(19)-triene ›Formula (I)--A=(A-2), R=H, R.sup.1 =R.sup.2 =CH.sub.3
CH.sub.2, R.sup.3 =.alpha.-CH.sub.3, R.sup.4 =R.sup.5 =H,
Y=(CH.sub.2).sub.4 !
The title compound is prepared by substituting ethyl lithium for methyl
lithium in (b) above and continuing the remainder of the procedure.
f)
25-Amino-1.alpha.,3.beta.-dihydroxy-24,26,26,26,27,27,27-heptakis-homo-9,1
0-secocholesta-5(Z),7,10(19)-triene ›Formula (I)--A=(A-2), R=H, R.sup.1
=R.sup.2 =CH.sub.3 (CH.sub.2).sub.3, R.sup.3 =.alpha.-CH.sub.3, R.sup.4
=R.sup.5 =H, Y=(CH.sub.2).sub.4 !
The title compound is prepared by substituting butyl lithium for methyl
lithium in (b) above and continuing the remainder of the procedure.
g)
25-Amino-1.alpha.,3.beta.,23-trihydroxy-9,10-secocholesta-5(Z),7,10(19)-tr
iene ›Formula (I)--A=(A-2), R=H, R.sup.1 =R.sup.2 =CH.sub.3, R.sup.3
=.alpha.-CH.sub.3, R.sup.4 =R.sup.5 =H, Y=CH.sub.2 CHOHCH.sub.2 !
The title compound is prepared by substituting
1.alpha.,3-bis-triisopropylsilyloxy-23-nor-9,10-secopregna-5(E),7,10(19)-t
riene-24-carboxaldehyde ›Formula (VII)--A=(A-3), R.sup.3 =.alpha.-CH.sub.3,
R.sup.4 =R.sup.5 =(i-Pr).sub.3 Si, Y.sup.b =CH.sub.2 ! for the bromoethyl
compound in (a) above and containing the remainder of the procedure.
25-Amino-lx,30-bis-triisopropylsilyloxy-24,26,27-tris-homo-9,10-secocholest
a-5(E),7,10(19),22,24(24a)-pentaene ›formula (I)--A=(A-3), R=H, R.sup.1
=R.sup.2 =CH.sub.3 CH.sub.2, R.sup.3 =.alpha.-CH.sub.3, R.sup.4 =R.sup.5
=(i-Pr).sub.3 Si, Y=CH.dbd.CH--CH.dbd.CH! is prepared by substituting
ethyl lithium for the methyl lithium and
1.alpha.,3.beta.-bis-triisopropylsilyloxy-24-cyano-24-homo-9,10-secochola-
5(E),7,10(19),22(E),24(E)-pentaene ›Formula (II)--A=(A-3), R.sup.1
=.alpha.-CH.sub.3, R.sup.4 =R.sup.5 (i-Pr).sub.3 Si,
Y=CH.dbd.CH--CH.dbd.CH! (Preparation 9 (b)) for the nitrile in (b) above.
Isomerisation and desilylation afford
25-amino-1.alpha.,3.beta.-dihydroxy-24,26,27-tris-homo-9,10-secocholesta-5
(Z),7,10(19), 22,24(24a)-pentaene ›Formula (I)--A=(A-2), R=H, R.sup.1
=R.sup.2 =CH.sub.3 CH.sub.2, R.sup.3 =.alpha.-CH.sub.3, R.sup.4 =R.sup.5
=OH, Y=CH.dbd.CH--CH.dbd.CH!.
EXAMPLE 6
25-Acetamido-1.alpha.,3.beta.-bis-triisopropylsilyloxy-2A-homo-9,10-secocho
lesta-5(Z),7,10(19)-triene ›Formula (I)--A=(A-2), R=CH.sub.3 CO, R.sup.1
=R.sup.2 =CH.sub.3, R.sup.3 =.alpha.-CH.sub.3, R.sup.4 =R.sup.5
=(i-Pr).sub.3 Si, Y=(CH.sub.2).sub.4 !
The 25-amino compound from Example 5 (c) above (48 mg) was treated with
acetic anhydride (0.026 ml) in pyridine (0.26 ml) and methylene chloride
(0.8 ml). After 2 hours storage at room temperature the reaction mixture
was cooled, treated with aqueous sodium bicarbonate and stirred for 2
hours, following which the product was extracted into ethyl acetate.
Chromatography gave title compound (24 mg). UV (Et.sub.2 O)
.lambda..sub.max 262-3, .lambda..sub.min 225 nm; IR .nu..sub.max
(CCl.sub.4) 3350, 3220 (NH), 1680 cm.sup.-1 (CONH); NMR (CCl.sub.4)
.delta. 0.53 (s, 18-H's), 1.8 (s, OCCH.sub.3), 4.1-4.46 (bm, 1,3-H's),
4.73, 5.06, 5.23 (each s, 19-H's, N--H), 5.66-6.36 (ABq, 6,7-H's)
b)
25-Acetamido-1.alpha.,3.beta.-dihydroxy-24-homo-9,10-secocholesta-5(Z),7,1
0(19)-triene ›Formula (I)--A=(A-2), R=CH.sub.3 CO, R.sup.1 =R.sup.2
=CH.sub.3, R.sup.3 =.alpha.-CH.sub.3, R.sup.4 =R.sup.5 =H,
Y=(CH.sub.2).sub.4 !
The silyl ether from (a) above (25 mg) was desilylated by treatment with
tetrabutylammonium fluoride (0.030 ml) in tetrahydrofuran (0.30 ml) at
room temperature for 4.5 hours. Chromatography gave the title compound (22
mg). UV (EtOH) .lambda..sub.max 263, .lambda..sub.min 225-6 nm; IR
.nu..sub.max (CHCl.sub.3) 3420, 3600 (NH, OH), 1670 cm3.sup.1 (CONH); NMR
(CDCl.sub.3) .delta. 0.56 (s, 18-H's), 0.86, 0.93 (d, 21-H's), 1.3 (d,
CH.sub.3'), 1.8 (s, OCCH.sub.3), 4.03-4.4 (bm, 1,3-H's), 4.9, 5.1, 5.23
(each s, 19-H's, N--H), 5.8-6.36 (ABq, 6,7-H's).
25-Benzamido-1.alpha.,3.beta.-bis-
trilsocropylsilyloxy-24-homo-9,10-secocholesta-5(Z),7,10(19)-triene
›formula (I)--A=-(A-2), R=C.sub.6 H.sub.5 CO, R.sup.1 =R.sup.2 =CH.sub.3,
R.sup.3 =.alpha.-CH.sub.3, R.sup.4 =R.sup.5 =(i-Pr).sub.3 Si,
Y=(CH.sub.2).sub.4 ! is prepared by use of benzoyl chloride and minor
modification (Schotten-Baumann) of the procedure of (a) above and may be
converted by desilylation into
25-benzamido-let,30-dihydroxy-24-homo-9,10-secocholesta-5(Z),7,10(19)-trie
ne ›formula (I)--A=(A-2), R=C.sub.6 H.sub.5 CO, R.sup.1 =R.sup.2 =CH.sub.3,
R.sup.3 =.alpha.-CH.sub.3, R.sup.4 =R.sup.5 =H, Y=(CH.sub.2).sub.4 !.
EXAMPLE 7
a)
25-Amino-1.alpha.,3.beta.-bis-triisopropylsilyloxy-26,27-bis-homo-9,10-sec
ocholesta-5(E),7,10(19)-trien-23-yne ›Formula (I)--A=(A-3), R=H, R.sup.1
=R.sup.2 =CH.sub.3:2, R.sup.3 =.alpha.-CH.sub.3 R.sup.4 =R.sup.5
=(i-Pr).sub.3 Si, Y=CH.sub.2 --C.tbd.C!
A solution of 3-amino-3-ethyl-1-pentyne ›Formula (VI)--R=H, R.sup.1
=R.sup.2 =CH.sub.3 CH.sub.2, n=O! (1.05 ml) in hexane (10.5 ml) containing
hexamethylphosphoramide (0.9 ml) was treated at 0.degree. with butyl
lithium (4.5 ml of a 1.4M solution in hexane), then kept with stirring at
5.degree. for 30 minutes, followed by 1.25 hours at room temperature. All
but 2.5 mMole of the solution (presumed to contain 6 mMole of acetylide
anion) was expelled. The remaining 2.5 mMole of anion was treated with
1.alpha.,3.beta.-bis-triisopropylsily-loxy-20.alpha.-bromomethyl-9,10-seco
pregna-5(E),7,10(19)-triene ›Formula (V)--A=(A-3), R.sup.3
=.alpha.-CH.sub.3 R.sup.4 =R.sup.5 =(i-Pr).sub.3 Si, L=Br, Y.sup.a
=CH.sub.2 ! (200 mg) and the resulting mixture stirred at 35.degree. for
21 hours. The reaction mixture was then cooled, treated with aqueous
ammonium chloride, and the product was extracted into ether.
Chromatography gave the title compound (68 mg) UV (Et.sub.2 O)
.lambda..sub.max 267, .lambda..sub.min 226 nm; NMR (CDCl.sub.4) .delta.
0.5 (s, 18-H's), 4.83 (S, 19-H's), 5.6-6.36 (ABq, 6,7-H's).
b)
25-Amino-1.alpha.,3.beta.-bis-triisopropylsilyloxy-26,27-bis-homo-9,10-sec
ocholesta-5(Z), 7,10(19)-trien-23-yne ›Formula (I)--A=(A-2), R=H, R.sup.1
=R.sup.2 =CH.sub.3 CH.sub.2, R.sup.3 =.alpha.-CH.sub.3 R.sup.4 =R.sup.5
=(i-Pr).sub.3 Si, Y=CH.sub.2 --C.tbd.C!
The product from (a) above (95 mg) was photoisomerized by irradiation for
30 minutes in solution in benzene (14 ml) containing phenazine (50 mg).
Chromatography gave the title compound (66.5 mg). UV (Et.sub.2 O)
.lambda..sub.max 260-1, .lambda..sub.min 226 nm; NMR (CCl.sub.4) .delta.
0.50 (S, 18-H's), 4.1-4.43 (bm, 1,3-H's), 4.73, 5.03 (each s, 19-H's),
5.9-6.23 (ABq, 6,7-H's).
c)
25-Amino-1.alpha.,3.beta.-dihydroxy-26,27-bis-homo-9,10-secocholesta-5(Z),
7,10(19)-trien-23-yne ›Formula (I)--A=(A-2), R=H R.sup.1 =R.sup.2 =CH.sub.3
CH.sub.2, R.sup.3 =.alpha.-CH.sub.3 R.sup.4 =R.sup.5 =H, Y=CH.sub.3
--C.tbd.C!
The silyl ether from (b) above (67 mg) was desilylated by treatment with
tetrabutylammonium fluoride (0.5 ml of a 1M solution in tetrahydrofuran)
in tetrahydrofuran (0.5 ml) at room temperature overnight. The product was
extracted into chloroform, washed with water and isolated by
chromatography to give the title compound (29 mg). UV (EtOH)
.lambda..sub.max 262-3, .lambda..sub.min 225 nm; IR (CHCl.sub.3)
.nu..sub.max 3200-3500, 3600 cm-.sup.-1 (OH, NH); NMR (CDCl.sub.3) .delta.
0.53 (s, 18-H's), 0.86-1.23 (m, 21-H's and Me-H's of Et's), 1.9 (m,
N--H's, exchanges with D.sub.2 O), 1.36-1.56 (m, Et-H's), 4.03-4.4 (bm,
1,3-H's), 4.86, 5.2 (ea. s, 19-H's), 5.76-6.33 (ABq, 6,7-H's).
By substituting
1.alpha.,3.beta.-bis-triisopropylsilyloxy-20.beta.-bromoethyl-9,10-secopre
gna-5(E),7,10(19)-triene ›Formula (V)--A=(A-3), R.sup.3 =.beta.-CH.sub.3,
R.sup.4 =R.sup.5 =(i-Pr).sub.3 Si, L=Br, Y=(CH.sub.2).sub.2 ! for the
steroid starting material in (a) above and following the remainder of the
procedure one may prepare
25-amino-1.alpha.,3.beta.-dihydroxy-20-epi-24,26,27-tris-homo-9,10-secocho
lesta-5(Z),7,10(19)-trien-24(24a)-yne ›Formula (I)--A=(A-2), R=H, R.sup.1
=R.sup.2 =CH.sub.3 CH.sub.2, R.sup.3 =.beta.-CH.sub.3, R.sup.4 =R.sup.5
=H, Y=(CH.sub.2).sub.2 --C.tbd.C--!.
By substituting 2-amino-2-methyl-4-pentyne ›Formula (VI)--R=H, R.sup.1
=R.sup.2 =CH.sub.3, n=1! for the alkyne in (a) above and following the
remaining procedures one obtains
25-amino-1.alpha.,3.beta.-dihydroxy-24-homo-9,10-secocholesta-5(Z),7,10(19
)-trien-24(24a)-yne ›Formula (1) A=(A-2), R=H, R.sup.1 =R.sup.2
=CH.sub.3,R.sup.3 =.alpha.-CH.sub.3, R.sup.4 =R.sup.5 =H, Y=CH.sub.2
--C.tbd.C--CH.sub.2 !.
EXAMPLE 8
Conversion of alkynylenyl side-chain to alkylenyl
The triene system of any of the steroid alkynes prepared acording to
Example 7, preferably having 5(E) configuration, is protected from
hydrogenation by reaction with a diazo dienophile (preferably phthalazine
dione) to form a Diels Alder adduct (between the 6- and 19-positions).
(GB-A-2114570). The resulting adduct (about 100 mg) in a mixture of
ethanol (5 ml) and benzene (5 ml) containing fresh platinum on charcoal
(5%, 100 mg) and sodium bicarbonate (50 mg) is kept under an atmosphere of
hydrogen until 2 molecular equivalents of hydrogen have been consumed
(about 20 hours). Filtration through Celite and removal of the solvents
gives substantially pure product. The phthalizine group is then removed as
described in the aforementioned British patent and the resulting 5(E)
vitamin may be photoisomerised and the silyl groups removed as in Example
7. In this fashion one may obtain
25-amino-1.alpha.,3.beta.-dihydroxy-20-epi-24,26,27-tris-homo-9,10-secocho
lesta-5(Z),7,10(19)-triene ›Formula (I) A=(A-2),R=H, R.sup.1 =R.sup.2
=CH.sub.3 CH.sub.2, R.sup.3 =.beta.-CH.sub.3, R.sup.4 =R.sup.5 -=H,
Y=(CH.sub.2).sub.4 ! from product of Example 7 (a).
EXAMPLE 9
a)
25-Amino-1.alpha.,3.beta.-bis-triisopropylsilyloxy-22-hydroxy-26,27-bis-ho
mo-9,10-secocholesta-5(E),7,10(19)-trien-23-yne ›Formula (I)--A=(A-3), R=H,
R.sup.1 =R.sup.2 =CH.sub.3 CH.sub.2, R.sup.3 =.alpha.CH.sub.3, R.sup.4
=R.sup.5 =(i-Pr).sub.3 Si, Y=CHOH--C.tbd.C!
A solution of 3-amino-3-ethyl-1-pentyne ›Formula (VI)--R=H, R.sup.1
=R.sup.2 =CH.sub.3 CH.sub.2, n=0! (0.78 ml) in hexane (6.5 ml) containing
hexamethylphosphoramide (0.6 ml) was treated at 5.degree. with butyl
lithium (3 ml of a 1.4M solution in hexane), then kept with stirring at
5.degree. for 30 minutes, followed by 1.25 hours at room temperature. A
portion of the solution (1/5, presumed to contain about 0.75 mMole of
acetylide anion) was added at -78.degree. to a solution of
1.alpha.,3.beta.-bis-triisopropylsilyloxy-20.alpha.-formyl-9,10-secopregna
-5(E),7,10(19)-triene ›Formula (VII)--A=(A-3), R.sup.3 =.alpha.-CH.sub.3,
R.sup.4 =R.sup.5 =(i-Pr).sub.3 Si, Y.sup.b =valence bond! (100 mg) and the
resulting mixture stirred while the temperature was allowed slowly to rise
to 200 (by which time the aldehyde had been consumed). The reaction
mixture was then cooled to -78.degree., treated with aqueous ammonium
chloride, and the product was extracted into ether. Chromatography gave
the title compound (94 mg). TV (Et.sub.2 O) .lambda..sub.max 268-9,
.lambda..sub.min 227 nm; NMR (CCl.sub.4) .delta. 0.55 (s, 18-H's),
4.23-4.56 (bm, 1,3-H's), 4.83 (s, 19-H's), 5.6-6.33 (ABq, 6,7-H's).
b)
25-Amino-1.alpha.,3.beta.-bis-triisopropylsilyloxy-22-hydroxy-26,27-bis-ho
mo-9,10-secocholesta-5(Z),7,10(19)-trien-23-yne ›Formula (I)--A=(A-2), R=H,
R.sup.1 =R.sup.2 =CH.sub.3 CH.sub.2, R.sup.3 =.alpha.-CH.sub.3, R.sup.4
=R.sup.5 =(i-Pr).sub.3 Si, Y=CHOH--C.tbd.C!
The product from (a) above (94 mg) was photoisomerized by irradiation for
40 minutes in solution in benzene (11 ml) containing phenazine (40 mg).
Chromatography gave the title compound (68 mg). UV (Et.sub.2 O)
.lambda..sub.max 261-2, .lambda..sub.min 225 nm; NMR (CCl.sub.4) .delta.
0.53 (s, 18-H's), 4.16-4.5 (bm, 1,3-H's), 4.73, 5.1 (each s, 19-H's),
5.73-6.06 (ABq, 6,7-H's).
c)
25-Amino-1.alpha.,3.beta.,22-trihydroxy-26,27-bis-homo-9,10-secocholesta-5
(Z),7,10(19)-trien-23-yne ›Formula (I)--A=(A-2), R=H, R.sup.1 =R.sup.2
=CH.sub.3 CH.sub.2, R.sup.3 =.alpha.-CH.sub.3, R.sup.4 =R.sup.5 =H.
Y=CHOH--C.tbd.C!
The silyl ether from (b) above (68 mg) was desilylated by treatment with
tetrabutylammonium fluoride (0.5 ml of a 1M solution in tetrahydrofuran)
in tetrahydrofuran (0.5 ml) at room temperature for 16 hours. TLC showed
some starting material so a further portion of tetrabutylammonium fluoride
(0.4 ml) was added and the reaction allowed to continue for a further 2
hours. The product was extracted into chloroform, washed with water and
isolated by chromatography (2.times.) followed by partition between
methylene chloride and water to give the title compound (18 mg). UV (EtOH)
.lambda..sub.max 263-4, .lambda..sub.min 225-6 nm; IR (CDCl.sub.3)
.nu..sub.max 3250-3450, 3600 cm.sup.-1 (OH, NH); NMR (CDCl.sub.3) .delta.
0.53 (s, 18-H's), 0.83-1.23 (m, 21-H's and Me-H's of Et's), 1.83 (m,
N--H's?), 1.36-1.56 (m, Et-H's), 4.1-4.5 (bm, 1,3-H's), 4.9, 5.23 (ea. s,
19-H's), 5.83-6.36 (ABq, 6,7-H's).
By substituting
1.alpha.,3.beta.-bis-triisopropylsilyloxy-23-nor-9,10-secopregna-5(E),7,10
(19)-triene-24-carboxaldehyde ›Formula (VII)--A=(A-3), R.sup.3
=.alpha.-CH.sub.3, R.sup.4 =R.sup.5 =(i-Pr).sub.3 Si, Y.sup.b =CH.sub.2 !
in (a) above and following the subsequent procedure one may prepare
25-amino-1a,3p,23-trihydroxy-24,26,27-tris-homo-9,10-secocholesta-5(Z),7,1
0(19)-triene-24(24a)-yne ›Formula (I)--A=(A-2), R=H, R.sup.1 =R.sup.2
=CH.sub.3 CH.sub.2, R.sup.3 =.alpha.-CH.sub.3, R.sup.4 =R.sup.5 =H,
Y=CH.sub.2 CHOH--C.tbd.C!.
By substituting 1,1-dimethylpropargyl amine ›Formula (VI)--R=H, R.sup.1
=R.sup.2 =CH.sub.3, n=0! for the aminoalkyne in (a) above and following
the subsequent procedures one may prepare
25-amino-1.alpha.,3.beta.,22-trihydroxy-9,10-secocholesta-5(Z),7,10(19)-tr
ien-23-yne ›Formula (I)--A=(A-2), R=H, R.sup.1 =R.sup.2 =CH.sub.3, R.sup.3
=.alpha.-CH.sub.3, R.sup.4 =R.sup.5 =H, Y=CHOH--C.tbd.C!.
d)
25-Amino-1.alpha.,3.beta.-dihydroxy-22-methoxy-26,27-bis-homo-9,10-secocho
lesta-5(Z),7,10(19)-trien-23-yne ›Formula (I)--A=(A-2), R=H, R.sup.1
=R.sup.2 =CH.sub.3 CH.sub.2, R.sup.3 =.alpha.-CH.sub.3 R.sup.4 =R.sup.5
=H, Y=CH(OCH.sub.3)--C.tbd.C!
The title compound is prepared by methylation of
25-amino-1.alpha.,3.beta.-bis-triisopropylsilyloxy-22-hydroxy-26,27-bis-ho
mo-9,10-secocholesta-5(Z),7,10(19)-trien-23-yne ›Formula (I)--A=(A-2), R=H,
R.sup.1 =R.sup.2 =CH.sub.3 CH.sub.2, R.sup.3 =.alpha.-CH.sub.3, R.sup.4
=R.sup.5 =(i-Pr).sub.3 Si, Y=CHOH--C.tbd.C! from (b) above using a
substantial excess of potassium t-butoxide in benzene containing
18-crown-6, followed by careful addition of methyl iodide (TLC control)
and then removing the silyl groups as in (c) above.
25-Amino-1.alpha.,3.beta.-dihydroxy-22-ethoxy-26,27-bis-homo-9,10-secochole
sta-5(Z),7,10(19)-trien-23-yne ›Formula (I)--A=(A-2), R=H, R.sup.1 =R.sup.2
=CH.sub.3 CH.sub.2, R.sup.3 =.alpha.-CH.sub.3 R.sup.4 =R.sup.5 =H,
Y=CH(OCH.sub.2 CH.sub.3)--C.tbd.C! and
25-amino-1.alpha.,3.beta.-dihydroxy-22-propoxy-26,27-bis-homo-9,10-secocho
lesta-5(Z),7,10(19)-trien-23-yne (Formula (I)--A=(A-2), R=H, R.sup.1
=R.sup.2 =CH.sub.3 CH.sub.2, R.sup.3 =.alpha.-CH.sub.3 R.sup.4 =R.sup.5
=H, Y=CH(OCH.sub.2 CH.sub.2 CH.sub.3)--C.tbd.C! are prepared as in (d)
above by replacement of methyl iodide by ethyl or propyl iodides
respectively.
EXAMPLE 10
a)
N-ethyl-25-amino-1.alpha.,3.beta.-bis-triisopropylsilyloxy-24-homo-9,10-se
cocholesta-5(Z),7,10(19)-triene ›Formula (I) --A=(A-2), R=CH.sub.3
CH.sub.2, R.sup.1 =R.sup.2 =CH.sub.3, R.sup.3 =.alpha.-CH.sub.3, R.sup.4
=R.sup.5 =(i-pr).sub.3 Si, L=Br, Y=CCH.sub.2).sub.4 !
A solution of
25-acetamido-l1,3p-bis-triisopropylsily-loxy-24-homo-9,10-secocholesta-5(Z
), 7,10(19)-triene ›Formula (I)--A=(A-2), R=CH.sub.3 CO, R.sup.1 =R.sup.2
=CH.sub.3, R.sup.3 =.alpha.-CH.sub.3, R.sup.4 =R.sup.5 =(i-Pr).sub.3 Si,
Y=(CH.sub.2).sub.4 ! prepared as in Example 6 (a) (44 mg) in
tetrahydrofuran (1.5 ml) containing lithium aluminium hydride (35 mg) is
heated under reflux with stirring until the starting material is consumed
(about 2.5 hours by TLC analysis), then cooled and treated with a few
drops of water. The reaction mixture is treated with sodium sulphate,
diluted with ether, and the etheral layer is decanted. Evaporation of the
solvent affords the title compound.
b)
N-ethyl-25-amino-1.alpha.,3.beta.-dihydroxy-24-homo-9,10-secocholesta-5(Z)
,7,10(19)-triene ›Formula (I)--A=(A-2), R=CH.sub.3 CH.sub.2, R.sup.1
=R.sup.2 =CH.sub.3, R.sup.3 =.alpha.-CH.sub.3, R.sup.4 =R.sup.5 -=H,
Y=(CH.sub.2).sub.4 !
The title compound is obtained by desilylation of the product of (a) above
in similar manner to Example 1 (d).
N-propyl-25-amino-1.alpha.,3.beta.-dihydroxy-24-homo-9,10-secocholesta-5(Z)
,7,10(19)-triene ›Formula (I)--A=(A-2), R=CH.sub.3 CH.sub.2 CH.sub.2,
R.sup.1 =R.sup.2 =CH.sub.3, R.sup.3 =.alpha.-CH.sub.3, R.sup.4 =R.sup.5
=H, Y=(CH.sub.2).sub.4 ! is similarly prepared by substituting
25-propionamido-1.alpha.,3.beta.-bis-triisopropylsilyloxy-24-homo-9,10-sec
ocholesta-5(Z),7,10(19)-triene ›Formula (I)--A=(A-2), R=CH.sub.3 CH.sub.2
CO, R.sup.1 =R.sup.2 =CH.sub.3, R.sup.3 =.alpha.-CH.sub.3, R.sup.4
=R.sup.5 =(i-Pr).sub.3 Si, Y=(CH.sub.2).sub.4 ! (prepared by substituting
propionyl chloride in Example 6 and increasing the reaction time to 4
hours) in (a) above and following the remainder of the procedure.
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